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Biomedical subjects

R R Anderson

Publications and source records attributed to R R Anderson.

At least 127 records · Page 7Linked to original sources

Light and electron microscopic analysis of tattoos treated by Q-switched ruby laser.

Short-pulse laser exposures can be used to alter pigmented structures in tissue by selective photothermolysis. Potential mechanisms of human tattoo pigment lightening with Q-switched ruby laser were explored by light and electron microscopy. Significant variation existed between and within tattoos. Electron microscopy of untreated tattoos revealed membrane-bound pigment granules, predominantly within fibroblasts and macrophages, and occasionally in mast cells. These granules contained pigment particles ranging from 2-in diameter. Immediately after exposure, dose-related injury was observed in cells containing pigment. Some pigment particles were smaller and lamellated. At fluences greater than or equal to 3 J/cm2, dermal vacuoles and homogenization of collagen bundles immediately adjacent to extracellular pigment were occasionally observed. A brisk neutrophilic infiltrate was apparent by 24 h. Eleven days later, the pigment was again intracellular. Half of the biopsies at 150 d revealed a mild persistent lymphocytic infiltrate. There was no fibrosis except for one case of clinical scarring. These findings confirm that short-pulse radiation can be used to selectively disrupt cells containing tattoo pigments. The physial alteration of pigment granules, redistribution, and elimination appear to account for clinical lightening of the tattoos.

Biopsy↗

Comparative studies of femtosecond to microsecond laser pulses on selective pigmented cell injury in skin.

Threshold radiant exposures for grossly apparent immediate whitening and ultrastructural alterations of melanosomes in black guinea pig skin were determined for a series of red visible laser pulses ranging from 4 x 10(-4) to 6.5 x 10(-14) s. Threshold exposures for melanosomal injury were found to be independent of pulsewidth when the pulsewidths were below the estimated thermal relaxation time of melanosomes. Threshold radiant exposures for melanosomal injury were found to increase when the pulsewidths were approximately equal to or above the thermal relaxation time of melanosomes. At longer pulse durations, fracturing of melanosomes was not observed despite the longer exposures necessary for injury. Instead, perimelanosomal vacuoles were noted. These findings are consistent with the theory of selective photothermolysis and provide evidence for the thermal initiation of melanosomal disruption.

Animals↗

Q-switched ruby laser irradiation of normal human skin. Histologic and ultrastructural findings.

The Q-switched ruby laser is used for treatment of tatoos. The effects of Q-switched ruby laser pulses on sun-exposed and sun-protected human skin, as well as senile lentigines, were investigated with clinical observation, light microscopy, and transmission electron microscopy. A pinpricklike sensation occurred at radiant exposures as low as 0.2 J/cm2. Immediate erythema, delayed edema, and immediate whitening occurred with increasing radiant exposure. The threshold for immediate whitening varied inversely with skin pigmentation, ranging from a mean of 1.4 J/cm2 in lentigines to 3.1 J/cm2 in sun-protected skin. Transmission electron microscopy showed immediate alteration of mature melanosomes and nuclei within keratinocytes and melanocytes, but stage I and II melanosomes were unaffected. Histologically, immediate injury was confined to the epidermis. There was minimal inflammatory response 1 day after exposure. After 1 week, subthreshold exposures induced hyperpigmentation, with epidermal hyperplasia and increased melanin staining noted histologically. At higher radiant exposures, hypopigmentation occurred with desquamation of a pigmented scale/crust. All sites returned to normal skin color and texture without scarring within 3 to 6 months. These observations suggest that the human skin response to selective photothermolysis of pigmented cells is similar to that reported in animal models, including low radiant exposure stimulation of melanogenesis and high radiant exposure lethal injury to pigmented epidermal cells.

Adult↗

Efficiency of opaque photoprotective agents in the visible light range.

"Opaque" physical sunscreens are important for photoprotection of individuals with visible light and UV-A photosensitivity such as those with porphyria, drug photoallergy, and polymorphous light eruption. Diffuse spectral transmittance of various thicknesses of opaque sunscreen formulations were measured from 350- to 800-nm range using a spectrophotometer equipped with an integrating sphere. Transmission through 20% zinc oxide paste was high and decreased minimally despite large increases in the sunscreen layer thickness. Adding a visible light absorber such as iron oxide to scattering sunscreens, however, substantially lowered transmittance below that predicted by the product of the transmittances for each component alone. Opaque sunscreens protected against hematoporphyrin derivative photosensitization of albino guinea pig skin; these results were quantitatively consistent with the in vitro findings. Poor photoprotection against visible light is obtained with white paste sunscreens, even if thick layers are applied. The addition of pigments to such sunscreens, however, greatly enhances photoprotection and cosmetic acceptability.

Animals↗

Polarized light examination and photography of the skin.

Light reflected from skin has two components: regular reflectance, or "glare" arising from the surface, and light backscattered from within the tissue. The regular reflectance contains the visual cues related to surface texture, whereas the backscattered component contains the cues related to pigmentation, erythema, infiltrates, vessels, and other intracutaneous structures. Unlike the backscattered component, regular reflectance preserves the plane of polarization of polarized incident light. Thus, viewing skin through a linear polarizer, under linearly polarized illumination, separates the two components of tissue reflectance. Thirty patients were examined and photographed in this manner. When the planes of polarization are parallel, images with enhanced surface detail are obtained. When the planes are orthogonal, wrinkles and surface detail disappear, and an enhanced view of vasculature and pigmented lesions is obtained. Simple, clinically useful techniques are presented.

Humans↗

Irradiation of pigmented melanoma cells with high intensity pulsed radiation generates acoustic waves and kills cells.

Photokilling of pigmented mouse melanoma cells (B-16) was investigated using pulsed high intensity visible radiation. Melanin acts as an endogenous chromophore, and 694 nm radiation with 40 nsec pulse duration and 0.5-3 X 10(7)w/cm2 intensity causes cell death. Irradiation of non-pigmented human melanoma cells (U1) or human squamous carcinoma cells (FaDu) under similar conditions did not kill the cells. Also, irradiation of B-16 cells with 300 microsec laser pulses (10(3)W/cm2) or with continuous wave (CW) radiation (10(-3)W/cm2) did not kill the cells. These data indicate that pigmented cell killing is due to absorption of radiation by melanin and that the pulsewidth and intensity of radiation play important roles in cell killing. The generation of acoustic waves due to absorption of the pulsed radiation by pigmented cells and by isolated melanosomes was demonstrated at 532 and 625 nm and 8.5 nsec pulse duration (10(7)-10(8) W/cm2); the amplitudes of the acoustic signals were approximately 2.5-3.0-fold higher at 532 nm compared with 625 nm, and they increased with increasing fluence. In contrast, irradiation of U1 or FaDu cells with comparable fluences and intensities did not generate acoustic waves. A possible correlation between the generation of photoacoustic waves and pigment cell death is proposed. Since the thermal relaxation time of melanosomes is 0.5-1.0 microsec, the mechanism proposed is that thermal confinement of high intensity, short-pulse visible radiation generates acoustic waves by thermal expansion, leading to mechanical damage to the cells.

Acoustics↗

Effect of blood upon the selective ablation of atherosclerotic plaque with a pulsed dye laser.

Laser angioplasty systems with laser energy preferentially absorbed by atherosclerotic plaque may offer a safe method of plaque removal. This study evaluated the effect of blood upon selective energy absorption using a pulsed dye laser at 480 nm. Intra-arterial laser irradiation of normal rabbit femoral arteries demonstrated a perforation threshold energy with blood perfusion of 13.1 mJ per pulse compared to 87.9 mJ with saline (P less than .0001), indicating a deleterious effect in the presence of blood. An adverse effect upon arterial healing at 3 days was noted in sheep following intra-arterial irradiation during blood but not saline perfusion. Normal and atherosclerotic human aorta ablation thresholds differed significantly (P less than .0002) under saline (plaque: 20 mJ and normal: 120 mJ) but the difference under blood (plaque: 5 mJ and normal: 20 mJ) was not significant. We conclude that absorption of laser energy by blood can reduce the effect of differential absorption by endogenous chromophores in normal and pathologic vascular tissues and, therefore, removal of blood may be a prerequisite for selective ablation of atherosclerotic plaques.

Animals↗

Thermotolerance and the heat shock response in normal human keratinocytes in culture.

Protective responses of normal human epidermal keratinocytes in culture, after exposure to elevated temperatures ("heat shock"), were examined. Cell viability, measured 24-48 h after a 20-min heat challenge at temperatures between 37 degrees C and 54 degrees C, declined sharply within a narrow 2 degrees-3 degrees C range. However, conditioning with a mild thermal pretreatment (40 degrees C or 42 degrees C for 1 h) protected the keratinocytes against a subsequent heat challenge. This induced thermotolerance was apparent when cells were challenged at 1, 3, and 6 h after the thermal pre-treatment, but disappeared by 24 h. Heating conditions that induce thermotolerance also stimulated the synthesis of heat-shock proteins (hsp) in these cells. Inductions of prominent 35S-methionine labeled bands at 70, 78, and 90 kDa were observed. However, the increases in synthesis of these heat-shock proteins did not correlate well with thermotolerance, because large increases were also observed at certain elevated temperatures that did not produce improved survival. Keratins observed in these cells (50 and 58 kDa classes) were not induced by heat shock. The development of thermotolerance, and the induction of hsp, were both completely blocked by 3'-deoxyadenosine (cordycepin), an inhibitor of newly synthesized messenger RNA, but not by adenosine, the normal analog. While heat-inducible mRNA apparently mediate some function important for the development of thermotolerance, the nature of that role remains speculative. Overall, our findings establish the existence of a functional thermal protective mechanism in human keratinocytes that appears to require the synthesis of new mRNA.

Cell Survival↗

Trace elements in milk of guinea pigs during a 20-day lactation.

This study was designed to determine patterns of change in milk trace elements over a complete lactation. Guinea pigs were selected because of ease of milking and short lactation duration. Common short-hair albino guinea pigs were milked using a milking apparatus for 4 to 6 h every other day following isolation from their offspring. Forty-eight samples of 4 ml each were analyzed for trace minerals using inductively coupled plasma spectroscopy. Observations represented 14 d scattered over 20 d of lactation. Eight elements with sufficiently high quantities to measure in milk were zinc (4.18 ppm), strontium (1.12), aluminum (.81), boron (.79), iron (.71), copper (.56), barium (.23), and manganese (.019). Regression analyses were used on rectilinear, quadratic, exponential and cubic models. Quadratic functions fit the data best in six elements, whereas in manganese the best model was the rectilinear. Indices of determination (R2) relating to day of lactation varied from R2 = .18 in boron to a high of .62 in strontium. For six elements, concentrations increased as milk volume decreased. The exception was zinc, which decreased in the latter half of lactation.

Aluminum↗

Treatment of tattoos by Q-switched ruby laser. A dose-response study.

Tattoo treatment with Q-switched ruby laser pulses (694 nm, 40 to 80 nanoseconds) was studied by clinical assessment and light and electron microscopy. Fifty-seven blue-black tattoos or portions thereof (35 amateur and 22 professional) were irradiated with 1.5 to 8.0 J/cm2 at a mean interval of 3 weeks. Substantial lightening or total clearing occurred in 18 (78%) of 23 amateur tattoos and 3 (23%) of 13 professional tattoos in which the protocol was completed. Response was related to exposure dose. Scarring occurred in one case, and persistent confettilike hypopigmentation was frequent. Optimal fluence was 4 to 8 J/cm2. Clinicohistologic correlation was poor. Q-switched ruby laser pulses can provide an effective treatment for tattoos.

Adult↗

Visible action spectrum for melanin-specific selective photothermolysis.

The skin of black and albino guinea pigs was irradiated with single, 750 nsec-long laser pulses at 435, 488, 530, and 560 nm in order to determine an action spectrum for the gross threshold response of immediate epidermal whitening. In addition, the immediate and delayed gross and histologic changes induced at, above, and below the threshold radiant exposures at all four wavelengths were studied. The action spectrum in the black guinea pigs was consistent with the reported absorption spectrum of DOPA-melanin. Histologically, there was epidermal damage immediately after radiant exposures at and above threshold at all four wavelengths. In addition, radiant exposures greater than threshold caused an immediate decrease in stainable epidermal pigment that was most marked at 435 and 488 nm. The healing response was also wavelength- and dose-dependent. Seven days after above-threshold exposures, there was little epidermal pigment in the 435 nm specimens. As wavelength increased, there was progressively more pigment, and in the 560 nm specimens, the epidermal pigment was equivalent to that seen in nonirradiated black guinea pig control specimens. Seven days after subthreshold radiant exposures, there was increased epidermal pigmentation and melanocytes at all four wavelengths. This was the most pronounced in the 435 nm specimens. There was no observable epidermal damage in albino guinea pig skin.

Animals↗

Selective photothermolysis of cutaneous pigmentation by Q-switched Nd: YAG laser pulses at 1064, 532, and 355 nm.

Exposure of skin to nanosecond-domain laser pulses affects the pigmentary system by a process called selective photothermolysis, in which melanosomes and pigmented cells are preferentially altered. Due to the broad absorption spectrum of melanin, this effect may occur with wavelengths that penetrate to vastly different depths within tissue, potentially producing different biologic responses. The effects of single near-ultraviolet (355 nm), visible (532 nm), and near infrared (1064 nm) pulses of 10-12 nsec duration were determined in guinea pig skin using gross, histologic, and electron microscopic observations. Threshold response in pigmented skin was a transient immediate ash-white discoloration, requiring 0.11, 0.20, and 1.0 J/cm2, at 355, 532, and 1064 nm, respectively. At each wavelength, melanosomes were ruptured within keratinocytes and melanocytes, with cytoplasmic and nuclear alterations. Delayed epidermal depigmentation occurred, followed by gradual repigmentation. Deep follicular cells were altered only at 532 and 1064 nm, which produced permanent leukotrichia. The action spectrum for threshold response was consistent with mechanisms implied by selective photothermolysis. These data may be useful for consideration of treatment for cutaneous pigmentation abnormalities or unwanted follicular pigmentation, or both.

Animals↗

Physiologic role of relaxin on mammary gland growth in rats.

The role of relaxin in stimulating growth of the mammary gland was assessed in ovariectomized and intact male rats for a period of 20 days. In addition to relaxin alone, the ovarian mammogenic hormones estradiol and progesterone were used in combination with relaxin and with each other to evaluate responses of mammae. Indices for mammary growth included wet weight, dry fat-free tissue, DNA, RNA, total protein, and collagen. Quantitative estimates of DNA and collagen represented the best indicators of parenchymal and stromal growth, respectively. Because changes in body weights were significantly different among hormonally administered groups, these were included as well. In Ovariectomized young rats, relaxin alone and in combination with estradiol and progesterone increased all indices significantly (P less than 0.01). The collagenous portion of total protein was high for the group receiving relaxin alone (62%) compared with the control group (46%). Relaxin administered along with estradiol and progesterone increased collagen accumulation to 73%, compared with 54% in the estradiol + progesterone group. Relaxin did not significantly increase growth indices when administered to male rats at 10 and 20 micrograms/day, while 30 micrograms stimulated a significant increase in total protein (P less than 0.05), suggesting that 30 micrograms of relaxin/day may be considered the basal concentration needed to induce a physiologic response in males. Relaxin induced a growth effect on mammae by synergizing with progesterone and estradiol in order to stimulate parenchymal proliferation, as noted by a DNA increase, and to increase stromal distensibility of the mammary pad by invoking accumulation of collagen and total protein in substituting for mammary adipose tissue.

Animals↗

Effect of xylazine in heifers under thermoneutral or heat stress conditions.

A study was performed to assess the effect of xylazine HCl (0.1 mg/kg of body weight, IV) in heifers maintained at thermoneutrality (18 C, 42% humidity) or under heat stress (33 C, 63% humidity) conditions. Xylazine caused 50 and 70% decreases in serum insulin concentrations in the thermoneutral and heat-stressed heifers, respectively. Xylazine-induced hypoinsulinemia was associated with hyperglycemia. In the thermoneutral group, serum glucose concentrations increased from a basal concentration of 75 mg/dl to 150 mg/dl after 15 minutes. In the heat stress group, the serum glucose concentration increased from 65 mg/dl to 105 mg/dl. Hyperglycemia peaked at 2 hours and remained high for 6 hours after xylazine administration. Heat-stressed heifers took a longer time (107 minutes) to stand than did heifers under thermoneutral conditions (41 minutes). The time to regain sensation to pain was significantly prolonged in heat-stressed heifers. Xylazine had no effect on body temperature and respiration rate in heifers under the thermoneutral condition, whereas it markedly induced hyperthermia and suppressed respiration rate in the heat-stressed heifers. Furthermore, the pulse rate was slightly decreased in thermoneutral heifers and was markedly decreased in the heat-stressed heifers.

Animals↗

Pigmented guinea pig skin irradiated with Q-switched ruby laser pulses. Morphologic and histologic findings.

Q-switched ruby laser pulses cause selective damage to cutaneous pigmented cells. Repair of this selective damage has not been well described. Therefore, using epilated pigmented and albino guinea pig skin, we studied the acute injury and tissue repair caused by 40-ns, Q-switched ruby laser pulses. Gross observation and light and electron microscopy were performed. No specific changes were evident in the albino guinea pigs. In pigmented animals, with radiant exposures of 0.4 J/cm2 or greater, white spots confined to the 2.5-mm exposure sites developed immediately and faded over 20 minutes. Delayed depigmentation occurred at seven to ten days, followed by full repigmentation by four to eight weeks. Regrowing hairs in sites irradiated at and above 0.4 J/cm2 remained white for at least four months. Histologically, vacuolation of pigment-laden cells was seen immediately in the epidermis and the follicular epithelium at exposures of 0.3 J/cm2 and greater. Melanosomal disruption was seen immediately by electron microscopy at and above 0.3 J/cm2. Over the next seven days, epidermal necrosis was followed by regeneration of a depigmented epidermis. By four months, melanosomes and melanin pigmentation had returned; however, hair follicles remained depigmented and devoid of melanocytes. This study demonstrates that selective melanosomal disruption caused by Q-switched ruby laser pulses leads to transient cutaneous depigmentation and persistent follicular depigmentation. Potential exists for selective treatment of pigmented epidermal and dermal lesions with this modality.

Animals↗

Ablation of rabbit liver, stomach, and colon with a pulsed holmium laser.

A pulsed holmium laser (wavelength 2.1 microns, pulse duration 250 microseconds) was used to ablate rabbit liver, stomach, and colon in vivo. Microscopic examination of the tissues revealed zones of thermal damage extending 0.5-1.0 mm from ablation sites. In addition, ablation rates were measured using a mass loss technique and found to increase linearly with delivered radiant exposure. The threshold radiant exposure for ablation was calculated to be 50 J/cm2 with a heat of ablation of 7000 J/cm3. Because the holmium laser produces less thermal necrosis than current endoscopic laser systems, such as the continuous-wave neodymium:YAG laser, and because the ablation rate can be precisely controlled, the holmium laser shows promise as an alternative method for endoscopic removal of tissue.

Animals↗