Metabolic Flexibility in Response to Within-Season Temperature Variability in House Sparrows.

The climatic variability hypothesis (CVH) posits that more flexible phenotypes should provide a fitness advantage for organisms experiencing more variable climates. While typically applied across geographically separated populations, whether this principle applies across seasons or other conditions (e.g., open vs. sheltered habitats) which differ in climatic variability remains essentially unstudied. In north-temperate climates, climatic variability in winter usually exceeds that in summer, so extending the CVH to within-population seasonal variation predicts that winter phenotypes should be more flexible than summer phenotypes. We tested this prediction of the within-season extension of the CVH by acclimating summer and winter-collected house sparrows (Passer domesticus) to 24, 5, and -10°C and measuring basal metabolic rate (BMR) and summit metabolic rate (Msum = maximum cold-induced metabolic rate) before and after acclimation (Accl). To examine mechanistic bases for metabolic variation, we measured flight muscle and heart masses and citrate synthase and ß-hydroxyacyl coA-dehydrogenase activities. BMR and Msum were higher for cold-acclimated than for warm-acclimated birds, and BMR was higher in winter than in summer birds. Contrary to our hypothesis of greater responses to cold Accl in winter birds, metabolic rates generally decreased over the Accl period for winter birds at all temperatures but increased at cold temperatures for summer birds. Flight muscle and heart masses were not significantly correlated with season or Accl treatment, except for supracoracoideus mass, which was lower at -10°C in winter, but flight muscle and heart masses were positively correlated with BMR and flight muscle mass was positively correlated with Msum. Catabolic enzyme activities were not clearly related to metabolic variation. Thus, our data suggest that predictions of the CVH may not be relevant when extended to seasonal temperature variability at the within-population scale. Indeed, these data suggest that metabolic rates are more prominently upregulated in summer than in winter in response to cold. Metabolic rates tended to decrease during Accl at all temperatures in winter, suggesting that initial metabolic rates at capture (higher in winter) influence metabolic Accl for captive birds.

Optical transfer diagnosis differentiating benign and malignant pigmented lesions in a simulated primary care practice.

BACKGROUND: The detection of melanoma poses a substantial challenge, particularly for primary care providers (PCPs) who may have limited training in discriminating between suspicious and benign melanocytic lesions. The noninvasive optical transfer diagnosis (OTD) method was designed to be used by PCPs in their decision-making process. OBJECTIVES: To assess the potential of the OTD method by developing, training and validating an OTD indication algorithm for automated discrimination between benign melanocytic lesions and malignant lesions, based on a set of 712 lesions. METHODS: The authors performed in vivoOTD capture and subsequent analysis of 712 pigmented lesions. Of the lesions, 415 were clinically and dermoscopically benign and 297 were dermoscopically suspicious or equivocal. After image capture, all suspicious or equivocal lesions were biopsied and examined histopathologically. RESULTS: Of the 297 suspicious or equivocal lesions, histopathological findings revealed 80 to be malignant (64 melanomas, 13 basal cell carcinomas and 3 squamous cell carcinomas). OTD misdiagnosed one of the 80 malignant lesions as benign (sensitivity, 99%). OTD specificity was 93% for the dermoscopically benign lesions, 73% for all lesions included in the study and 36% for the clinically suspicious but histopathologically benign lesions. CONCLUSIONS: High sensitivity and specificity, as provided by OTD in this preliminary study, would help PCPs reduce the number of referrals for dermatology consultation, excision or biopsy. Further studies are planned for screening patients in a primary care setting, with comparisons of OTD results with biopsy or dermoscopy results.

Educational and practice gaps in the management of volar melanocytic lesions.

BACKGROUND: The benign and malignant patterns of acral melanocytic naevi (AMN) and acral melanomas (AM) have been defined in a series of retrospective studies. A three-step algorithm was developed to determine when to biopsy acral melanocytic lesions. This algorithm has only been applied to a Japanese population. OBJECTIVES: Our study aimed to review the current management strategy of acral melanocytic lesions and to investigate the utility of the three-step algorithm in a predominately Caucasian cohort. METHODS: A retrospective search of the pathology and image databases at Mayo Clinic was performed between the years 2006 and 2016. Only cases located on a volar surface with dermoscopic images were included. Two dermatologists reviewed all dermoscopic images and assigned a global dermoscopic pattern. Clinical and follow-up data were gathered by chart review. All lesions with known diameter and pathological diagnosis were used for the three-step algorithm. RESULTS: Regular fibrillar and ridge patterns were more likely to be biopsied (P = 0.01). The majority of AMN (58.1%) and AM (60%) biopsied were due to physician-deemed concerning dermoscopic patterns. 39.2% of these cases were parallel furrow, lattice-like or regular fibrillar. When patients were asked to follow-up within a 3- to 6-month period, only 16.7% of the patients returned within that interval. The three-step algorithm would have correctly identified four of five AM for biopsy, missing a 6 mm, multicomponent, invasive melanoma. CONCLUSION: We found one major educational gap in the recognition of low-risk lesions with high rates of biopsy of the fibrillary pattern. Recognizing low-risk dermoscopic patterns could reduce the rate of biopsy of AMN by 23.3%. We identified two major practice gaps, poor patient compliance with follow-up and the potential insensitivity of the three-step algorithm to small multicomponent acral melanocytic lesions.

Comparative Transcriptomics of Seasonal Phenotypic Flexibility in Two North American Songbirds.

Phenotypic flexibility allows organisms to reversibly alter their phenotypes to match the changing demands of seasonal environments. Because phenotypic flexibility is mediated, at least in part, by changes in gene regulation, comparative transcriptomic studies can provide insights into the mechanistic underpinnings of seasonal phenotypic flexibility, and the extent to which regulatory responses to changing seasons are conserved across species. To begin to address these questions, we sampled individuals of two resident North American songbird species, American goldfinch (Spinus tristis) and black-capped chickadee (Poecile atricapillus) in summer and winter to measure seasonal variation in pectoralis transcriptomic profiles and to identify conserved and species-specific elements of these seasonal profiles. We found that very few genes exhibited divergent responses to changes in season between species, and instead, a core set of over 1200 genes responded to season concordantly in both species. Moreover, several key metabolic pathways, regulatory networks, and gene functional classes were commonly recruited to induce seasonal phenotypic shifts in these species. The seasonal transcriptomic responses mirror winter increases in pectoralis mass and cellular metabolic intensity documented in previous studies of both species, suggesting that these seasonal phenotypic responses are due in part to changes in gene expression. Despite growing evidence of muscle nonshivering thermogenesis (NST) in young precocial birds, we did not find strong evidence of upregulation of genes putatively involved in NST during winter in either species, suggesting that seasonal modification of muscular NST is not a prominent contributor to winter increases in thermogenic capacity for adult passerine birds. Together, these results provide the first comprehensive overview of potential common regulatory mechanisms underlying seasonally flexible phenotypes in wild, free-ranging birds.

The diagnostic impact of limited, screening obstetric ultrasound when performed by midwives in rural Uganda.

OBJECTIVE: To evaluate the diagnostic impact of limited obstetric ultrasound (US) in identifying high-risk pregnancies when used as a screening tool by midwives in rural Uganda. STUDY DESIGN: This was an institutional review board-approved prospective study of expecting mothers in rural Uganda who underwent clinical and US exams as part of their standard antenatal care visit in a local health center in the Isingiro district of Uganda. The midwives documented clinical impressions before performing a limited obstetric US on the same patient. The clinical findings were then compared with the subsequent US findings to determine the diagnostic impact. The midwives were US-naive before participating in the 6-week training course for limited obstetric US. RESULT: Midwife-performed screening obstetric US altered the clinical diagnosis in up to 12% clinical encounters. This diagnostic impact is less (6.7 to 7.4%) if the early third trimester diagnosis of malpresentation is excluded. The quality assurance review of midwives' imaging demonstrated 100% sensitivity and specificity in the diagnosing gestational number, and 90% sensitivity and 96% specificity in the diagnosis of fetal presentation. CONCLUSION: Limited, screening obstetric US performed by midwives with focused, obstetric US training demonstrates the diagnostic impact for identifying conditions associated with high-risk pregnancies in 6.7 to 12% of patients screened. The limited obstetric US improved diagnosis of early pregnancy complication as well as later gestation twins and malpresentation. Midwives who have undergone focused 6-week limited obstetric US training proved capable of diagnosing twins and fetal presentation with high sensitivity and specificity.

Are summit metabolism and thermogenic endurance correlated in winter-acclimatized passerine birds?

Small birds exhibiting marked winter improvement of cold tolerance also show elevated summit metabolic rates (maximum cold-induced metabolic rate) in winter relative to summer. However, relatively large increases in cold tolerance can occur with only minor increments of maximum cold-induced metabolic rate and geographic variation in cold tolerance is not always positively correlated with variation in maximum cold-induced metabolic rate. Thus, it is uncertain whether maximum cold-induced metabolic rate and cold tolerance are phenotypically correlated in small birds and no previous study has directly examined this relationship. I measured maximum cold-induced metabolic rate and cold tolerance (i.e., thermogenic endurance) over three winters in black-capped chickadees Poecile atricapillus, American tree sparrows Spizella arborea, and dark-eyed juncos Junco hyemalis. For raw thermogenic endurance data, residuals of maximum cold-induced metabolic rate and thermogenic endurance from mass regressions were significantly and positively correlated in juncos and tree sparrows, and their correlation approached significance for chickadees. Log10 transformation of thermogenic endurance and mass data gave similar results. These data provide the first direct evidence for a phenotypic correlation between maximum cold-induced metabolic rate and thermogenic endurance in small birds, although much of the variance in thermogenic endurance is explained by factors other than maximum cold-induced metabolic rate and the degree of correlation differs among species. Nevertheless, these data suggest that physiological adjustments producing elevated thermogenic endurance also produce elevated maximum cold-induced metabolic rate in small birds.

Time course for cryoprotectant synthesis in the freeze-tolerant chorus frog, Pseudacris triseriata.

Increases in liver glycogen phosphorylase activity, along with inhibition of glycogen synthetase and phosphofructokinase-1, are associated with elevated cryoprotectant (glucose) levels during freezing in some freeze-tolerant anurans. In contrast, freeze-tolerant chorus frogs, Pseudacris triseriata, accumulate glucose during freezing but exhibit no increase in phosphorylase activity following 24-h freezing bouts. In the present study, chorus frogs were frozen for 5- and 30-min and 2- and 24-h durations. After freezing, glucose, glycogen, and glycogen phosphorylase and synthetase activities were measured in leg muscle and liver to determine if enzyme activities varied over shorter freezing durations, along with glucose accumulation. Liver and muscle glucose levels rose significantly (5-12-fold) during freezing. Glycogen showed no significant temporal variation in liver, but in muscle, glycogen was significantly elevated after 24 h of freezing relative to 5 and 30 min-frozen treatments. Hepatic phosphorylase a and total phosphorylase activities, as well as the percent of the enzyme in the active form, showed no significant temporal variation following freezing. Muscle phosphorylase a activity and percent active form increased significantly after 24 h of freezing, suggesting some enhancement of enzyme function following freezing in muscle. However, the significance of this enhanced activity is uncertain because of the concurrent increase in muscle glycogen with freezing. Neither glucose 6-phosphate independent (I) nor total glycogen synthetase activities were reduced in liver or muscle during freezing. Thus, chorus frogs displayed typical cryoprotectant accumulation compared with other freeze-tolerant anurans, but freezing did not significantly alter activities of hepatic enzymes associated with glycogen metabolism.

Characteristics of veal calves upon arrival, at 28 and 84 days, and at end of the production cycle.

Holstein bull calves arriving at five special-fed veal farms (eight production groups) were scored for various physical condition traits and blood-sampled within 2 h after arrival and at 28 d, 84 d, and 1 wk prior to slaughter at 116 to 143 d. Of 1179 total calves in the production groups, 758 calves were scored and evaluated. Blood cell analyses (red and white blood cell counts, hemoglobin, and packed cell volume) were conducted at all four sampling times; total serum protein concentration was obtained at 0 and 28 d. The study was initiated in autumn and ended the following autumn. Mean initial and final body weights were 46.3+/-0.17 and 209.7+/-0.77 kg; mean mortality was 2.5%. Average daily gain of the eight groups ranged from 1.23 to 1.70 kg/d. Subjective scores of 5 = excellent to 1 = very poor condition were used to evaluate 16 different physical condition characteristics. With the exception of leg joint, hoof, and foot scores, most of the physical condition scores exhibited improvement during the first 28 d. Foot and leg impairments did not appear to hinder the ambulatory ability of the calves during the production period. Physical condition scores at d 0 and 28 were generally not related to numbers or types of medical treatments (enteric, respiratory, other, or total) or to average daily gain during the production period. Means for most erythrocytic and leukocytic traits upon arrival (d 0) were within normal ranges, although 27.4% of the calves were clinically or marginally anemic. Final mean hemoglobin and packed cell volume were 8.53 g/dl and 26.1%. Forty-three percent of the calves at d 0 were colostral deficient, assuming that total serum protein concentrations of <5.5 g/dl indicate colostral deficiency. No blood trait was consistently correlated with body weight gain when gain during the production period was divided into quartiles and the blood traits were averaged by gain quartile. Calves in the lowest serum total protein quartile (mean 4.58 g/dl) had more respiratory and total medical treatments than quartiles with higher total protein means. Dairy bull calves arriving at veal production units after transporting from the dairy farm to the auction market (or other collection facility) have several physical impairments. However, most of these physical impairments are improved early in the veal feeding period and are not generally related to subsequent growth rate or medical treatment.

Evidence for a proximate influence of winter temperature on metabolism in passerine birds.

The roles of ultimate and proximate factors in regulating basal and summit metabolic rates of passerine birds during winter have received little study, and the extent to which winter temperatures affect these variables is unknown. To address this question, we measured basal and summit (maximum cold-induced) metabolic rates in black-capped chickadees (Poecile atricapillus), dark-eyed juncos (Junco hyemalis), and American tree sparrows (Spizella arborea) during winters from 1991/1992 to 1997 in southeastern South Dakota. Both temperature and these metabolic rates varied within and among winters. Least-squares regression revealed significant negative relationships for normalized basal and summit metabolism against mean winter temperature for all species pooled (R2=0.62 to 0.69, P

Implant sequence effects in intact male Holstein veal calves: live and slaughter traits.

Seven sequences of growth promotant implants were used in special-fed intact male Holstein veal calves (n = 443). Calves received implants 4 d after arrival at the veal barn, 42, and 84. The following implants were used: placebo (0), Z (36 mg zeranol), ET (20 mg estradiol, 200 mg testosterone), EP/2 (10 mg estradiol, 100 mg progesterone), EP (20 mg estradiol, 200 mg progesterone), and EBA (24 mg estradiol, 120 mg trenbolone acetate). The following sequences were compared: 0-0-0 (negative control), 0-ET-ET, Z-ET-ET, 0-EP-EP, Z-EP-EP, 0-EP/2-EBA, and Z-0-EBA. From 0 to 42 d, Z implants increased (P<.05) ADG by 3.4% compared to placebo. However, implant schemes without an initial Z implant (0-ET-ET and 0-EP-EP) had higher (P<.05) mean ADG for the period from d 42 to 84. From 84 d to the end of the experiment, only the 0-EP/2-EBA treatment increased (P<.05) ADG compared to 0-0-0. Over the entire trial 0-ET-ET, 0-EP-EP, Z-EP-EP, and 0-EP/2-EBA implant sequences increased (P<.05) ADG by 3.2, 3.2, 2.4, and 4.7%, respectively, compared to the 0-0-0 sequence. Blood traits measured within 2 wk before slaughter were not affected by implant sequence, except that sequences with EP had higher (P<.05) leukocyte counts than were observed for the other sequences. Testicular weight was less (P<.01) for all of the implant sequences than for the negative control and less (P<.05) for Z-ET-ET than for 0-ET-ET, 0-EP-EP, 0-EP/2-EBA, and Z-0-EBA. The type and frequency of medical treatments did not differ among implant sequences for any of the 42-d phases, or over the entire trial. Generally, the growth promotant implants currently approved for beef cattle resulted in approximately 50% of the increase in growth rate in Holstein intact bull calves, as has been observed in beef-type steers or heifers.

Implant sequence effects in intact male Holstein veal calves: carcass characteristics.

Seven sequences of growth promotant implants were used in intact male Holstein veal calves (n = 443). Implants were administered on d 0 (within 4 d after arrival at the veal barn), 42, and 84. The implants used were placebo (0), Z (36 mg zeranol), ET (20 mg estradiol, 200 mg testosterone), EP/2 (10 mg estradiol, 100 mg progesterone), EP (20 mg estradiol, 200 mg progesterone), and EBA (24 mg estradiol, 120 mg trenbolone acetate). The following sequences were compared: 0-0-0 (negative control), 0-ET-ET, Z-ET-ET, 0-EP-EP, Z-EP-EP, 0-EP/2-EBA, and Z-0-EBA. Sequences 0-EP-EP, Z-EP-EP, and 0-EP/2-EBA increased (P<.05) carcass weight from 3.3 to 3.9% compared to nonimplanted controls. There were no differences (P>.05) in percentage of carcass weight accounted for by the fore vs. rear halves of carcasses, suggesting there was no difference in the distribution of weight. Although there were differences in longissimus area, the results were not consistent, except that there was a trend for longissimus area to be increased by the use of estrogenic-androgenic implants (ET and EBA). There were no differences among implant sequences for carcass conformation, fat cover, muscle texture, marbling/ feathering, muscle color, or muscle chemical composition. Of four implant sequences (0-0-0, 0-ET-ET, 0-EP-EP, and 0-EP/2-EBA) tested for differences in Warner-Bratzler shear force tenderness, the latter two sequences averaged higher (P<.05) for shear force than did the negative control. These results suggest that aggressive implant strategies in young, intact Holstein bull calves (raised as veal) have minimal effects on carcass characteristics.

Comparison of the 16S ribosomal DNA sequences from the intracellular agents of proliferative enteritis in a hamster, deer, and ostrich with the sequence of a porcine isolate of Lawsonia intracellularis.

Proliferative enteritis is an enteric disease that affects a variety of animals. The causative agent in swine has been determined to be an obligate intracellular bacterium, Lawsonia intracellularis, related to the sulfate-reducing bacterium Desulfovibrio desulfuricans. The intracellular agents found in the lesions of different animal species are antigenically similar. In addition, strains from the pig, ferret, and hamster have been shown to be genetically similar. In this study we performed a partial 16S ribosomal DNA sequence analysis on the intracellular agent of proliferative enteritis from a hamster, a deer, and an ostrich and compared these sequences to that of the porcine L. intracellularis isolate. Results of this study indicate that the intracellular agents from these species with proliferative enteritis have high sequence similarity, indicating that they are all in the genus Lawsonia and that they may also be the same species, L. intracellularis.

Diagnosis of proliferative enteritis in frozen and formalin-fixed, paraffin-embedded tissues from a hamster, horse, deer and ostrich using a Lawsonia intracellularis-specific multiplex PCR assay.

Proliferative enteritis (PE) is an enteric disease that has been reported in a variety of animals. It is caused by an obligate intracellular bacterium identified in swine as Lawsonia intracellularis. The organism can be detected ante-mortem in swine with PE using molecular diagnostic methods. The disease can be diagnosed post-mortem in all species by gross examination of tissues and special histologic staining procedures. In this study we extracted total DNA from frozen or formalin-fixed, paraffin-embedded tissues from cases of pig, hamster, horse, deer and ostrich PE. The samples were subjected to a multiplex PCR reaction using primers specific for a swine isolate of L. intracellularis. Identical sized PCR products were detected in samples from all animals with PE and the specificity of the PCR reaction for L. intracellularis was demonstrated by Southern-blotting and hybridization using specific probes. These results suggest that the intracellular organism of PE in these species are all very closely related to the causative agent of PE in swine, L. intracellularis. In addition, this multiplex PCR assay can be used to detect the organism in frozen or archival tissues, facilitating retrospective diagnosis of PE.

Freezing tolerance/intolerance and cryoprotectant synthesis in terrestrially overwintering anurans in the Great Plains, USA.

Mechanistic bases for freezing tolerance in anurans have been well-studied only in wood frogs, Rana sylvatica, so comprehensive explanations for the mechanisms and evolution of freezing tolerance in anurans are lacking. We measured crystallization temperatures, freezing tolerance/intolerance, and tissue glucose and glycogen phosphorylase activities in frozen and unfrozen winter-acclimated Pseudacris triseriata, Bufo cognatus and B. woodhousei. Freezing occurred at higher subzero temperatures on wet substrate than on dry substrate in all species, indicating susceptibility to inoculative freezing. P. triseriata was freeze-tolerant, but survival was dependent on the level of supercooling prior to freezing. All Bufo were freezing intolerant, regardless of crystallization temperature. Glucose was significantly elevated by freezing in both liver (35-fold) and leg muscle (22-fold) in winter P. triseriata, but only liver glucose was significantly elevated in B. cognatus. However, freezing did not alter glycogen phosphorylase activity in either species. Liver phosphorylase activity was significantly higher in P. triseriata than in B. cognatus, suggesting that capacity for mobilizing glucose from liver glycogen is associated with freezing tolerance. Summer measurements of liver phosphorylase activity, however, did not differ between species. Thus, P. triseriata, but not B. cognatus, exhibited winter increment of liver phosphorylase activity that is correlated with the development of freezing tolerance.

Colonization of rabbits by Pasteurella multocida: serum IgG responses following intranasal challenge with serologically distinct isolates.

Enzyme-linked immunosorbent assays (ELISAs) and immunoblots were used to measure serum IgG responses in rabbits which were intranasally challenged with Pasteurella multocida. The responses to two serologically distinct isolates (isolate 1, serotype 3:A and isolate 10, serotype 1:D) were compared and then correlated with the ability of the isolates to colonize the nasal passages. Five rabbits were challenged with each isolate (10(5) CFU); nasal washings and sera were collected weekly for 8 weeks. Serum IgG levels were measured by ELISA and immunoblots, using bacterial whole cells and lipopolysaccharides (LPSs) as antigens. The serum IgG response to isolate 1 was evident earlier and was significantly stronger than the response to isolate 10 (P less than 0.025). Immunoblots supported this observation and confirmed that both isolates elicited antibodies which reacted with bacterial protein and LPS antigens, with antibody to protein detectable before antibody to LPS. Results of weekly nasal cultures suggested that the antibody response data could be explained by a difference in the ability of the isolates to colonize the nasal passages: isolate 1 was recovered from four of five rabbits for 8 weeks, whereas isolate 10 was recovered for a maximum of 2 weeks, even when the challenge dose was increased tenfold. The strong response elicited by isolate 1 was therefore probably a result of persistent colonization, whereas the weak response to isolate 10 may have resulted from an inability to persistently colonize the nasal passages. The results of this study demonstrate that isolates of P. multocida elicit antibody responses of differing intensities and vary in their ability to colonize the nasal passages.(ABSTRACT TRUNCATED AT 250 WORDS)

Embolic abscesses in hereditary hemorrhagic telangiectasia.

Hereditary hemorrhagic telangiectasia is an inherited disorder in which patients may have multiple telangiectases and arteriovenous fistulas in the skin and internal organs. Patients can suffer from a variety of serious clinical complications, including abscess formation. We report two patients in whom neurologic symptoms developed from embolic abscesses, one for whom this complication was fatal. The reported incidence and microbiologic features of this complication are similar to that of endocarditis in patients with valvular heart disease. We believe that patients with hereditary hemorrhagic telangiectasia should receive similar antibacterial prophylaxis for procedures placing them at risk for bacteremia.

A dermatitis-eosinophilia syndrome. Treatment with methylprednisolone pulse therapy.

A 47-year-old man had a generalized, eczematous erythroderma and eosinophilia one week after a wasp sting. These changes persisted for four months despite intensive topical therapy and oral corticosteroids. He was then given corticosteroid pulse therapy with methylprednisolone sodium succinate (2 g, intravenously). One week later, a second pulse treatment was administered. This therapy was followed by permanent resolution of the dermatitis within two weeks.

Methylprednisolone inhibits pemphigus acantholysis in skin cultures.

Glucocorticosteroids are used to treat patients with pemphigus, but the mechanism of action is unknown. We studied the effect of methylprednisolone on acantholysis induced in vitro by incubation of normal skin with plasma from a patient with pemphigus. Normal human breast skin was maintained in organ cultures for several days in Ham F-10 medium. Plasma from a patient with active pemphigus vulgaris caused suprabasilar epidermal acantholysis when added to this culture system. In control cultures (F-10 medium and fetal bovine serum), no acantholysis occurred. Acantholysis was prevented when breast skin was preincubated for 24 h in a 0.25 mM solution of methylprednisolone in F-10 medium and fetal bovine serum, suggesting that methylprednisolone directly inhibits acantholysis. No suppression of acantholysis occurred when the methylprednisolone was added to the culture system simultaneously with the pemphigus plasma, suggesting a time requirement for alteration of cellular events. The inhibition of acantholysis was not caused by cell death since methylprednisolone did not alter keratinocyte viability as determined by exclusion of trypan blue dye when keratinocytes were exposed to pemphigus plasma. Similarly, the inhibition of acantholysis was not due to dissolution, alteration, or coating of pemphigus antigen on epidermal cells, since the intercellular antibodies in the plasma bound as well to methylprednisolone-treated epidermis as to untreated epidermis.