Search

Background Traumatic brain injury (TBI) is a leading cause of adult morbidity and mortality. Individuals with TBI have impairments in both cognitive and motor domains. Motor improvements post-TBI are attributable to adaptive neuroplasticity and motor learning. Majority of the studies focus on remediation of balance and mobility issues. There is limited understanding on the use of interventions for upper limb (UL) motor improvements in this population. Objective We examined the evidence regarding the effectiveness of different interventions to augment UL motor improvement after a TBI. Methods We systematically examined the evidence published in English from 1990–2020. The modified Downs and Black checklist helped assess study quality (total score: 28). Studies were classified as excellent: 24–28, good: 19–23, fair: 14–18, and poor: ≤13 in quality. Effect sizes helped quantify intervention effectiveness. Results Twenty-three studies were retrieved. Study quality was excellent (n = 1), good (n = 5) or fair (n = 17). Interventions used included strategies to decrease muscle tone (n = 6), constraint induced movement therapy (n = 4), virtual reality gaming (n = 5), non-invasive stimulation (n = 3), arm motor ability training (n = 1), stem cell transplant (n = 1), task-oriented training (n = 2), and feedback provision (n = 1). Motor impairment outcomes included Fugl-Meyer Assessment, Modified Ashworth Scale, and kinematic outcomes (error and movement straightness). Activity limitation outcomes included Wolf Motor Function Test and Motor Activity Log (MAL). Effect sizes for majority of the interventions ranged from medium (.5-.79) to large (≥.8). Only ten studies included retention testing. Conclusion There is preliminary evidence that using some interventions may enhance UL motor improvement after a TBI. Answers to emergent questions can help select the most appropriate interventions in this population.

(1) What are the structures indicated by the blue arrows?(2) In this silver stain, which step/reagent is omitted that would normally prevent the yellow background staining?(3) What disease often pr...

BackgroundTraumatic Brain Injury (TBI) is a leading cause of adult morbidity and mortality. Individuals with TBI have impairments in both cognitive and motor domains. Motor improvements post-TBI are attributable to adaptive neuroplasticity and motor learning. Majority of the studies focus on remediation of balance and mobility issues. There is limited understanding on the use of interventions for upper limb (UL) motor improvements in this population.ObjectiveWe examined the evidence regarding the effectiveness of different interventions to augment UL motor improvement after a TBI.MethodsWe systematically examined the evidence published in English from 1990-2020. The modified Downs and Black checklist helped assess study quality (total score:28). Studies were classified as excellent:24-28, good:19-23, fair:14-18 and poor:≤13 in quality. Effect sizes helped quantify intervention effectiveness.ResultsTwenty-three studies were retrieved. Study quality was excellent(n=1), good(n=5) or fair(n=17). Interventions used included strategies to decrease muscle tone (n=6), constraint induced movement therapy (n=4), virtual reality gaming (n=5), noninvasive stimulation (n=3), arm motor ability training (n=1), stem-cell transplant (n=1); task-oriented training (n=2) and feedback provision (n=1). Motor impairment outcomes included Fugl-Meyer Assessment, Modified Ashworth Scale, and kinematic outcomes (error and movement straightness). Activity limitation outcomes included Wolf Motor Function Test and Motor Activity Log. Effect sizes for majority of the interventions ranged from medium(0.5-0.79) to large(≥0.8). Only ten studies included retention testing.ConclusionThere is preliminary evidence that using some interventions may enhance UL motor improvement after a TBI. Answers to emergent questions can help select the most appropriate interventions in this population.

What is this stain?What are the pink/purple filamentous structures?What substance results in the positive pink stain? Chromic Acid-SchiffFungal hyphaeCarbohydrates in the walls of the fungal cell walls

Objectives The objective of the current study was to catalog hair shaft abnormalities in individuals with ectodermal dysplasia (ED) syndromes using scanning electron microscopy (SEM) and to compare the findings with those in unaffected controls. This is the second of a two-part study, the first of which used light microscopy as the modality and was previously published. Methods Scanning electron microscopy was performed in a blinded manner on hair shafts from 65 subjects with seven types of ED syndromes and 41 unaffected control subjects. Assessment was performed along the length of the shaft and in cross section. Setting Hair donations were collected at the 28th Annual National Family Conference held by the National Foundation for Ectodermal Dysplasia. Control subjects were recruited from a private dermatology practice and an academic children's hospital outpatient dermatology clinic. Results SEM identified various pathologic hair shaft abnormalities in each type of ED and in control patients. When hairs with all types of ED were grouped together and compared with those of control patients, the difference in the presence of small diameter and shallow and deep grooves was statistically significant (p

We have previously demonstrated that CD4+CD25+ regulatory T cells (Tregs) activated during the course of feline immunodeficiency virus (FIV) infection suppress CD8+ CTL function in a TGF-β–dependent fashion, inhibiting IFN-γ and IL-2 production and inducing G1 cell-cycle arrest. In this article, we describe the molecular events occurring at the IL-2 promoter leading to suppression of IL-2 production. These experiments demonstrate that Foxp3 induced by lentivirus-activated Tregs in the CD8+ target cells binds to the IL-2 promoter, actively repressing IL-2 transcription. We further demonstrate that the chronic activation of CD8+ T cells during FIV infection results in chromatin remodeling at the IL-2 promoter, specifically, demethylation of CpG residues. These DNA modifications occur during active transcription and translation of IL-2; however, these changes render the IL-2 promoter permissive to Foxp3-induced transcriptional repression. These data help explain, in part, the seemingly paradoxical observations that CD8+ T cells displaying an activation phenotype exhibit altered antiviral function. Further, we demonstrate that blocking demethylation of CpG residues at the IL-2 promoter inhibits Foxp3 binding, suggesting a potential mechanism for rescue and/or reactivation of CD8+ T cells. Using the FIV model for lentiviral persistence, these studies provide a framework for understanding how immune activation combined with Treg-mediated suppression may affect CD8+ T cell IL-2 transcription, maturation, and antiviral function.

A unique p38α MAPK–MK2 pathway inhibitor, CDD-450, is used to uncover the function of this protein complex in inflammasome priming signals. Importantly, CDD-450 is as efficacious as global p38α MAPK inhibitors in decreasing inflammation in disease models., p38α activation of multiple effectors may underlie the failure of global p38α inhibitors in clinical trials. A unique inhibitor (CDD-450) was developed that selectively blocked p38α activation of the proinflammatory kinase MK2 while sparing p38α activation of PRAK and ATF2. Next, the hypothesis that the p38α–MK2 complex mediates inflammasome priming cues was tested. CDD-450 had no effect on NLRP3 expression, but it decreased IL-1β expression by promoting IL-1β mRNA degradation. Thus, IL-1β is regulated not only transcriptionally by NF-κB and posttranslationally by the inflammasomes but also posttranscriptionally by p38α–MK2. CDD-450 also accelerated TNF-α and IL-6 mRNA decay, inhibited inflammation in mice with cryopyrinopathy, and was as efficacious as global p38α inhibitors in attenuating arthritis in rats and cytokine expression by cells from patients with cryopyrinopathy and rheumatoid arthritis. These findings have clinical translation implications as CDD-450 offers the potential to avoid tachyphylaxis associated with global p38α inhibitors that may result from their inhibition of non-MK2 substrates involved in antiinflammatory and housekeeping responses.

Vandetanib (ZD6474, Zactima) is a novel inhibitor of multiple tyrosine kinases that has been used for the treatment of various malignancies including medullary thyroid carcinoma.1 The drug functions primarily via inhibitory effects on vascular endothelial growth factor receptors, epidermal growth factor receptors, and the RET-tyrosine kinase system.2 As with other targeted chemotherapy agents, several cutaneous side effects including folliculitis, palmar-plantar hyperkeratosis, desquamation, cutaneous hyperpigmentation, and photosensitization have been reported.3, 4 Photosensitivity, although less commonly reported, was found to make up 37% of cutaneous adverse effects in one case series.3 Most of these patients were found to exhibit a sunburnlike erythema in sun-exposed regions with subsequent desquamation. However, a small subset of patients exhibited a phenotypically lichenoid photodermatitis and a lymphocytic interface dermatitis with basal vacuolization and keratinocyte apoptosis upon histopathology. Only a handful of cases describing similar lichenoid eruptions after vandetanib use have been reported, most of which developed within a few weeks after exposure.5, 6, 7, 8 We present a patient with a late-onset lichenoid photodermatitis that began 5 months after initiation of vandetanib therapy.

Diabetic non-healing wounds are a major clinical problem. The mechanisms leading to poor wound healing in diabetes are multifactorial but unresolved inflammation may be a major contributing factor. The complement system (CS) is the most potent inflammatory cascade in humans and contributes to poor wound healing in animal models. Signal transducer and activator of transcription 4 (STAT4) is a transcription factor expressed in immune and adipose cells and contributes to upregulation of some inflammatory chemokines and cytokines. Persistent CS and STAT4 expression in diabetic wounds may thus contribute to chronic inflammation and delayed healing. The purpose of this study was to characterize CS and STAT4 in early diabetic wounds using db/db mice as a diabetic skin wound model. The CS was found to be activated early in the diabetic wounds as demonstrated by increased anaphylatoxin C5a in wound fluid and C3-fragment deposition by immunostaining. These changes were associated with a 76% increase in nucleated cells in the wounds of db/db mice vs. controls. The novel classical CS inhibitor, Peptide Inhibitor of Complement C1 (PIC1) reduced inflammation when added directly or saturated in an acellular skin scaffold, as reflected by reduced CS components and leukocyte infiltration. A significant increase in expression of STAT4 and the downstream macrophage chemokine CCL2 and its receptor CCR2 were also found in the early wounds of db/db mice compared to non-diabetic controls. These studies provide evidence for two new promising targets to reduce unresolved inflammation and to improve healing of diabetic skin wounds.

Background Virtual microscopy is used for teaching medical students and residents and for in-training and certification examinations in the United States. However, no existing studies compare diagnostic accuracy using virtual slides and photomicrographs. The objective of this study was to compare diagnostic accuracy of dermatopathologists and pathologists using photomicrographs vs. digitized images, through a self-assessment examination, and to elucidate assessment of virtual dermatopathology. Methods Forty-five dermatopathologists and pathologists received a randomized combination of 15 virtual slides and photomicrographs with corresponding clinical photographs and information in a self-assessment examination format. Descriptive data analysis and comparison of groups were performed using a chi-square test. Results Diagnostic accuracy in dermatopathology using virtual dermatopathology or photomicrographs was similar: 0.70 vs. 0.73 respectively, p = 0.465. Order of administration of virtual dermatopathology and photomicrographs did not affect diagnostic accuracy. The vast majority (93%) of the participants felt the virtual slides were adequate for diagnosis and that virtual dermatopathology represented a useful tool for learning; 90% felt that virtual dermatopathology is useful tool for teaching dermatopathology. Conclusion No significant difference was observed in diagnostic accuracy using virtual dermatopathology compared to photomicrographs. Most participants felt virtual dermatopathology was adequate for diagnostic purposes and a useful tool for learning and teaching dermatopathology.

The incidence of primary cutaneous melanoma has been increasing dramatically for several decades. Melanoma accounts for the majority of skin cancer–related deaths, but treatment is nearly always curative with early detection of disease. In this update of the guidelines of care, we will discuss the treatment of patients with primary cutaneous melanoma. We will discuss biopsy techniques of a lesion clinically suspicious for melanoma and offer recommendations for the histopathologic interpretation of cutaneous melanoma. We will offer recommendations for the use of laboratory and imaging tests in the initial workup of patients with newly diagnosed melanoma and for follow-up of asymptomatic patients. With regard to treatment of primary cutaneous melanoma, we will provide recommendations for surgical margins and briefly discuss nonsurgical treatments. Finally, we will discuss the value and limitations of sentinel lymph node biopsy and offer recommendations for its use in patients with primary cutaneous melanoma.

Background: Virtual microscopy can be used to teach histology and pathology and for in-training and certification examinations. A few online consultation websites already utilize virtual microscopy, thereby expanding the role of telemedicine in dermatopathology. There are, however, relatively few studies comparing the diagnostic accuracy and acceptability of virtual slides compared to traditional glass slides. Methods: Ten Nordic dermatopathologists and pathologists were given a randomized combination of 20 virtual and glass slides and asked to identify the diagnoses. They were then asked to give their impressions about the virtual images. Descriptive data analysis and comparison of groups using Fisher's exact test were performed. Objective: To compare the diagnostic ability of dermatopathologists and pathologists in two image formats: the traditional (glass) microscopic slides, and whole mount digitized images, and to elucidate their assessment of virtual microscopy in dermatopathology. Results: Dermatopathologists and pathologists performed similarly in diagnosing dermatopathological disorders using virtual slides vs. glass slides, virtual 0.85 and glass 0.81, P=0.286. The order of administration of virtual or glass slides did not affect the percentage of questions answered correctly. Seven of nine participants completing the questionnaire, felt virtual microscopy is useful for both learning and testing. Conclusion: There was no significant difference in the participants' diagnostic ability using virtual slides compared with glass slides. Most participants agreed that virtual microscopy is a useful tool for learning and testing.

Up to 90% of the patients infected with human immunodeficiency virus are affected with a dermatologic disease, which may be of inflammatory, infectious, or neoplastic origin. This review focuses on the clinical and histological features of noninfectious inflammatory dermatoses of human immunodeficiency virus including psoriasis, pityriasis rubra pilaris, seborrheic dermatitis, eosinophilic folliculitis, pruritic papular eruption of human immunodeficiency virus, photodermatoses, eczematous dermatoses, and drug eruptions.

Activation of the p38 kinase pathway in immune cells leads to the transcriptional and translational regulation of proinflammatory cytokines. Mitogen-activated protein kinase-activated protein kinase 2 (MK2), a direct downstream substrate of p38 kinase, regulates lipopolysaccharide (LPS)-stimulated tumor necrosis factor alpha (TNFalpha) and interleukin-6 (IL-6) production through modulating the stability and translation of these mRNAs. Developing small-molecule inhibitors of MK2 may yield anti-inflammatory efficacy with a different safety profile relative to p38 kinase inhibitors. This article describes the pharmacologic properties of a benzothiophene MK2 inhibitor, PF-3644022 [(10R)-10-methyl-3-(6-methylpyridin-3-yl)-9,10,11,12-tetrahydro-8H-[1,4]diazepino[5',6':4,5]thieno[3,2-f]quinolin-8-one]. PF-3644022 is a potent freely reversible ATP-competitive compound that inhibits MK2 activity (K(i) = 3 nM) with good selectivity when profiled against 200 human kinases. In the human U937 monocytic cell line or peripheral blood mononuclear cells, PF-3644022 potently inhibits TNFalpha production with similar activity (IC(50) = 160 nM). PF-3644022 blocks TNFalpha and IL-6 production in LPS-stimulated human whole blood with IC(50) values of 1.6 and 10.3 microM, respectively. Inhibition of TNFalpha in U937 cells and blood correlates closely with inhibition of phospho-heat shock protein 27, a target biomarker of MK2 activity. PF-3644022 displays good pharmacokinetic parameters in rats and is orally efficacious in both the rat acute LPS-induced TNFalpha model and the chronic streptococcal cell wall-induced arthritis model. Dose-dependent inhibition of TNFalpha production in the acute model and inhibition of paw swelling in the chronic model is observed with ED(50) values of 6.9 and 20 mg/kg, respectively. PF-3644022 efficacy in the chronic inflammation model is strongly correlated with maintaining a C(min) higher than the EC(50) measured in the rat LPS-induced TNFalpha model.

Congress is grappling with ways to fund health care in the future. Much of the focus rests on paying physicians for their patients' outcomes, rather than the current system of payment for services provided during each visit. The years ahead will be years of change for American health care, with an increasing emphasis on the comparison of patient outcomes and measures of quality. Patient safety initiatives will be an integral part of the overall strategy to improve American health care. Part one of this two-part series on patient safety examines what we know about patient safety in dermatology, including data from medicolegal claims and published data on patient safety in the setting of office-based surgery. The article also focuses on how medical societies, payers, the US government, and the Board of Medical Specialties are responding to calls for accountability and improvements in patient safety. Learning objectives After completing this learning activity, participants should be able to identify risks to patient safety based on an understanding of the major causes of legal claims against dermatologists, use published patient safety data to improve the practice of office surgery, and be able to improve patient safety through an understanding of requirements for maintenance of certification.

Summary Virtual microscopy is being used in medical schools to teach histology and pathology. It is also being used in resident education, in-training examinations (dermatology), and certification examinations (pathology). There are, however, few studies comparing its diagnostic accuracy and acceptability compared with traditional glass slides. This study sought to compare residents' abilities in diagnosing dermatopathology disorders in 2 image formats (traditional microscopic slides and whole mount digitized images) and to assess their perceptions of virtual microscopy in dermatopathology. Residents in dermatology and pathology training programs at 14 institutions were given a randomized combination of 20 virtual and glass slides and were asked to identify the diagnoses from multiple foils. They were then asked to give their impressions about the virtual images. Descriptive data analysis and comparison of groups using Pearson χ 2 and Fisher exact tests for categorical variables and Student t test for continuous variables were performed. Residents in dermatology and pathology performed similarly in diagnosing dermatopathology disorders using virtual slides or glass slides (mean [SD] correct for virtual versus glass, 5.48 (1.72) versus 5.57 (2.06); P = .70). The order of administration of virtual versus glass slides did not affect the percentage of questions answered correctly. Most residents supported the use of virtual microscopy as a learning aid, whereas fewer favored its use in testing (79% versus 44%, respectively). Residents performed similarly in making dermatologic diagnoses using virtual slides compared with glass slides despite the residents' preference for the latter.

Food product development courses at The Pennsylvania State University and Saint Joseph's University were combined into a single course with a common faculty, syllabus, and web-based course management system. Industry specialists made weekly presentations on critical aspects of the product development and marketing process, via compressed video conferencing and the Web, to students at both locations. Cross-institutional interdisciplinary student teams completed a comprehensive plan for developing, evaluating, and launching a new product. Assessment of student learning revealed an enhanced understanding of all aspects of the product innovation, development, and marketing processes. The net result is a model learning system transcending disciplines, place, and time.

Spun soy fibers fractured by the tensile test, Warner-Bratzler shear, Kramer Shear Press and a modified Volodkevich wedge were examined with the scanning electron microscope. Fracture in the tensile test occurred almost always at a macrocavity, the weakest point in the fiber. Warner-Bratzler shear induced compression and flow at the zone of rupture, but not as markedly as in the case of the modified Volodkevich wedge. Fracture by the Kramer Shear Press was the closest to pure shear. Some of the exposed fracture surfaces revealed internal structures in the fibers such as macrocavities and distinct skin and core sections. The mechanism of rupture by these tests was more complex than expected.

Background Both increases and decreases in the incidence of subsequent malignancies in melanoma patients have been reported. We examined the database of the Indiana University Cancer Center to determine whether there is an association between malignant melanoma and noncutaneous malignancies. Objective We searched for evidence of noncutaneous malignancies in a cohort of melanoma patients. Methods Patients with microscopically confirmed malignant melanoma diagnosed between January 1987 and March 2001 were analyzed. This cohort was investigated for noncutaneous malignancies occurring either before or after the diagnosis of melanoma. The standardized incidence ratios (SIR) were calculated as the ratio of the observed to the expected number of patients with second malignancies, and 95% confidence intervals (95% CI) around the SIR were estimated from the cumulative Poisson distribution. Results A total of 955 patients with melanoma (498 males and 457 females) were documented over the 14-year period. Sixty-nine noncutaneous malignancies were identified in 59 (6.2%) melanoma patients (39 males and 20 females). There was a higher risk of non-Hodgkin's lymphoma (SIR = 1.91; 95% CI, 0.88–3.62) in men and renal cell carcinoma (SIR = 2.41, 95% CI, 0.97–4.97) in men. In female patients, however, there was no higher risk of noncutaneous malignancies. Conclusions This study did not show a higher risk of prostate cancer, gastrointestinal cancer, leukemia, endometrial cancer, or cancer of the nerve and neuroendocrine systems in melanoma patients. No female patients incurred a higher risk of noncutaneous cancers. The increased risk of non-Hodgkin's lymphoma and renal cell carcinoma in men might be attributed to a mutual carcinogenic exposure, an aberration of cell-mediated immunity, a shared genetic susceptibility, increased medical surveillance among cancer patients, a post-therapy effect after cancer management, or factors not as yet clear. Close monitoring of melanoma patients for signs of second malignancy is warranted.

Background: Various structural components of human skin biopsy specimens are difficult to visualize using conventional histologic approaches. Methods: We used two-photon microscopy and advanced imaging software to render three-dimensional (3D) images of in situ nerves, blood vessels, and hair follicles labeled with various fluorescent markers. Archived frozen human skin biopsy specimens were cryosectioned up to 150 µm in thickness and fluorescently stained with rhodamine- or fluorescein-labeled antibodies or lectins. Optical sections were collected by two-photon microscopy and the resulting data sets were analyzed in three dimensions using Voxx software. Results: Reconstructed image volumes demonstrated the complex 3D morphology of nerves, blood vessels and adnexal structures in normal mucocutaneous tissue. Conclusion: Two-photon microscopy and Voxx rendering software allow for detailed 3D visualization of structures within human mucocutaneous biopsy specimens, as they appear in situ, and facilitate objective interpretation of variations in their morphology. These techniques may be used to investigate disorders involving cutaneous structures that are difficult to visualize by means of traditional microscopy.

Myelopoietins (MPOs) are a family of recombinant chimeric proteins that are both interleukin-3 (IL-3) receptor and granulocyte colony-stimulating factor (G-CSF) receptor agonists. In this study, MPO molecules containing one of three different IL-3 receptor agonists linked with a common G-CSF receptor agonist have been examined for their IL-3 receptor binding characteristics. Binding to the alpha-subunit of the IL-3 receptor revealed that the affinity of the MPO molecules was 1.7-3.4-fold less potent than those of their individual cognate IL-3 receptor agonists. The affinity decrease was reflected in the MPO chimeras having approximately 2-fold slower dissociation rates and 2.7-5.5-fold slower association rates than the corresponding specific IL-3 receptor agonists alone. The affinity of binding of the MPO molecules to the heteromultimeric alphabeta IL-3 receptor expressed on TF-1 cells was either 3-, 10-, or 42-fold less potent than that of the individual cognate IL-3 receptor agonist. Biophysical data from nuclear magnetic resonance, near-UV circular dichroism, dynamic light scattering, analytical ultracentrifugation, and size exclusion chromatography experiments determined that there were significant tertiary structural differences between the MPO molecules. These structural differences suggested that the IL-3 and G-CSF receptor agonist domains within the MPO chimera may perturb one another to varying degrees. Thus, the differential modulation of affinity observed in IL-3 receptor binding may be a direct result of the magnitude of these interdomain interactions.

The lipooligosaccharide (LOS) of Haemophilus ducreyi contains a major glycoform that is immunochemically identical to paragloboside, a glycosphingolipid precursor of major human blood group antigens. We recently identified the gene responsible for the glucosyltransferase activity and constructed an isogenic mutant (35000glu-) deficient in this activity. 35000glu- makes an LOS that consists only of the heptose trisaccharide core and 2-keto-deoxyoctulosonic acid (KDO). For this study, the mutant was reconstructed in the 35000HP (human passaged [HP]) background. Five human subjects were inoculated with 35000HP and 35000HPglu- in a dose-response trial. The pustule formation rates were 40% (95% confidence interval [CI], 13.7 to 72.6%) at 10 sites for 35000HP and 46.7% (95% CI, 24.8 to 69.9%) at 15 sites for 35000HPglu-. The histopathology and recovery rates of H. ducreyi from surface cultures and biopsies obtained from mutant and parent sites were similar. These results indicate that the expression of glycoforms with sugar moieties extending beyond the heptose trisaccharide core is not required for pustule formation by H. ducreyi in humans.

Calciphylaxis or calcific uremic arteriolopathy (CUA) is a fatal disease in dialysis patients due to calcification of cutaneous blood vessels. The pathogenesis has been attributed to elevated parathyroid hormone (PTH). However, recent studies evaluating vascular calcification in nondialysis patients have found that the smooth muscle cells play an active role, including production of the bone matrix protein osteopontin. To examine the involvement of various clinical parameters and smooth muscle cells of CUA, we performed a case-control analysis comparing 10 CUA patients with our current dialysis patients. Available histologic sections were immunostained for osteopontin, markers of smooth muscle cells, endothelial cells, and macrophages. Compared with our current dialysis population, patients with CUA were more likely to be obese, white, and female (P < 0.02). Comparison of laboratory values found CUA patients with lower serum albumin, greater serum phosphorus, and greater calcium X phosphorus product (P < 0.01). In contrast, there was no difference in the concentration of PTH or calcium between the 2 groups. Immunostaining of calcified blood vessels showed that all calcified vessels stained positive for osteopontin, whereas all the noncalcifed vessels showed no osteopontin localization. Staining for smooth muscle alpha-actin decreased in the medial layer with calcification, with cells appearing to be sloughed off, leading to near occlusion of the vessel lumen. Our case-control study demonstrates that hyperphosphatemia and an elevated calcium X phosphorus product is associated with CUA. Histologic examination suggests that the calcification is associated with increased expression of osteopontin by smooth muscle cells.

Objective The progenipoietins, a class of engineered proteins containing both fetal liver tyrosine kinase-3 and granulocyte colony-stimulating factor receptor agonist activities, were functionally characterized in vitro and in vivo. Materials and Methods Four representative progenipoietins were evaluated for receptor binding, receptor-dependent cell proliferation, colony-forming unit activity, and their effects on hematopoiesis in the C57BL/6 mouse. Results The progenipoietins bound to fetal liver tyrosine kinase-3 and the granulocyte colony-stimulating factor receptor with affinities within twofold to threefold of the native ligands, and each progenipoietin bound simultaneously to both fetal liver tyrosine kinase-3 and the granulocyte colony-stimulating factor receptor. The progenipoietins exhibited different levels of activity in receptor-dependent cell proliferation assays. The fetal liver tyrosine kinase-3–dependent cell proliferation activity of three of four progenipoietins was decreased sixfold to 33-fold relative to native fetal liver tyrosine kinase-3 ligand, while granulocyte colony-stimulating factor receptor–dependent activity of the progenipoietins was within twofold to threefold of native granulocyte colony-stimulating factor. At nonsaturating concentrations, the progenipoietins stimulated colony formation to a greater extent than the equimolar combination of fetal liver tyrosine kinase-3 and granulocyte colony-stimulating factor. Treatment of mice with the progenipoietins yielded dramatic increases in peripheral blood and splenic white blood cells, polymorphonuclear leukocytes, and dendritic cells. Conclusion These preclinical results demonstrate that the progenipoietins are potent hematopoietic growth factors that stimulate cells in a receptor-dependent manner. When administered in vivo, the progenipoietins effectively promote the generation of multiple cell lineages. Thus, in both in vitro and in vivo settings, the progenipoietins as single molecules exhibit the synergistic activity of the combination of fetal liver tyrosine kinase-3 and granulocyte colony-stimulating factor.

Mutations in the NF1 tumor suppressor gene cause neurofibromatosis type I (NF1), a disease characterized by the formation of cutaneous neurofibromas infiltrated with a high density of degranulating mast cells. A hallmark of cell lines generated from NF1 patients or Nf1-deficient mice is their propensity to hyperproliferate. Neurofibromin, the protein encoded by NF1, negatively regulates p21(ras) activity by accelerating the conversion of Ras-GTP to Ras-GDP. However, identification of alterations in specific p21(ras) effector pathways that control proliferation in NF1-deficient cells is incomplete and critical for understanding disease pathogenesis. Recent studies have suggested that the proliferative effects of p21(ras) may depend on signaling outputs from the small Rho GTPases, Rac and Rho, but the physiologic importance of these interactions in an animal disease model has not been established. Using a genetic intercross between Nf1(+/)- and Rac2(-)(/)- mice, we now provide genetic evidence to support a biochemical model where hyperactivation of the extracellular signal-regulated kinase (ERK) via the hematopoietic-specific Rho GTPase, Rac2, directly contributes to the hyperproliferation of Nf1-deficient mast cells in vitro and in vivo. Further, we demonstrate that Rac2 functions as mediator of cross-talk between phosphoinositide 3-kinase (PI-3K) and the classical p21(ras)-Raf-Mek-ERK pathway to confer a distinct proliferative advantage to Nf1(+/)- mast cells. Thus, these studies identify Rac2 as a novel mediator of cross-talk between PI-3K and the p21(ras)-ERK pathway which functions to alter the cellular phenotype of a cell lineage involved in the pathologic complications of a common genetic disease.

Haemophilus ducreyiexpresses a peptidoglycan-associated lipoprotein (PAL) that exhibits extensive homology toHaemophilus influenzaeprotein 6. We constructed an isogenic PAL mutant (35000HP-SMS4) by the use of a suicide vector that containslacZas a counterselectable marker.H. ducreyi35000HP-SMS4 and its parent, 35000HP, had similar growth rates in broth and similar lipooligosaccharide profiles. 35000HP-SMS4 formed smaller, more transparent colonies than 35000HP and, unlike its parent, was hypersensitive to antibiotics. Complementation of the mutant intransrestored the parental phenotypes. To test whether expression of PAL is required for virulence, nine human volunteers were experimentally infected. Each subject was inoculated with two doses (41 to 89 CFU) of live 35000HP and one dose of heat-killed bacteria on one arm and with three doses (ranging from 28 to 800 CFU) of live 35000HP-SMS4 on the other arm. Papules developed at similar rates at sites inoculated with the mutant or parent but were significantly smaller at mutant-inoculated sites than at parent-inoculated sites. The pustule formation rate was 72% (95% confidence interval [CI], 46.5 to 90.3%) at 18 parent sites and 11% (95% CI, 2.4 to 29.2%) at 27 mutant sites (P< 0.0001). The rates of recovery ofH. ducreyifrom surface cultures were 8% (n= 130; 95% CI, 4.3 to 14.6%) for parent-inoculated sites and 0% (n= 120; 95% CI, 0.0 to 2.5%) for mutant-inoculated sites (P< 0.001).H. ducreyiwas recovered from six of seven biopsied parent-inoculated sites and from one of three biopsied mutant-inoculated sites. Confocal microscopy confirmed that the bacteria present in a mutant inoculation site pustule lacked a PAL-specific epitope. Although biosafety regulations precluded our testing the complemented mutant in humans, these results suggest that expression of PAL facilitates the ability ofH. ducreyito progress to the pustular stage of disease.

Background: Lobular capillary hemangiomas (pyogenic granulomas) occur on both mucosal and cutaneous surfaces. There are conflicting data regarding the increased prevalence of lobular capillary hemangiomas in female versus male subjects. Some studies have noted a female predominance of lobular capillary hemangiomas, but other studies do not reveal such a disparity. Because of an increased prevalence during pregnancy, oral tumors are also known as "granuloma gravidarum" or "pregnancy tumors." A hormonal influence for these mucosal lesions has been postulated. There are, however, no studies that address a possible relationship between hormones and cutaneous lesions. Objective: This study presents the epidemiology of lobular capillary hemangiomas, with an emphasis on cutaneous lesions. Methods: We reviewed 63,759 dermatopathology reports from a regional, private dermatopathology laboratory and found 325 cases of lobular capillary hemangiomas over a 1-year period. Results: In our study of lobular capillary hemangiomas, cutaneous lesions accounted for 86%, with mucosal lesions representing only 12% of cases. Seven cases were excluded (one was intravascular, two were subcutaneous, and in 4 the location was not specified). Overall, male patients outnumbered female patients. The peak incidence for cutaneous lobular capillary hemangiomas was found in the second decade of life. The most common cutaneous sites were the trunk, upper extremities, and head. Mucosal lesions were primarily seen on the lips, gingiva, and tongue, and these affected females more than males by a ratio of 2:1, most commonly in the fourth decade of life. Conclusion: Cutaneous lobular capillary hemangiomas were equally prevalent in male and female patients. This would refute a female hormonal influence in the induction of cutaneous lobular capillary hemangiomas. Our data may suggest a hormonal influence on mucosal lesions because mucosal lobular capillary hemangiomas were twice as common in female patients. However, the small number of lesions in our study precludes us from making such a conclusion. (J Am Acad Dermatol 2000;42:1012–6.)

Haemophilus ducreyi expresses 2 OmpA homologs, designated MOMP and OmpA2, whose genes are arranged in tandem on the chromosome. Northern blot analysis indicated that momp and ompA2 are transcribed independently. Sequences of the momp open reading frame (ORF) lacking the transcriptional start site were amplified by PCR, and an Ω-Km2 cassette was ligated into the ORF. A plasmid containing this construction was electroporated into H. ducreyi 35000HP, and an isogenic MOMP-deficient mutant (35000HP-SMS2) was generated by allele exchange. In Southern blotting, 35000HP-SMS2 contained one copy of the Ω-Km2 cassette in momp . 35000HP and 35000HP-SMS2 had similar outer membrane protein (OMP) and lipooligosaccharide profiles and growth rates except for up-regulation of a putative porin protein in the mutant. Five subjects were inoculated with three doses of live 35000HP-SMS2 on one arm and two doses of live 35000HP and one dose of a heat-killed control on the other arm in a double-blind escalating dose-response trial. Pustules developed at 7 of 10 sites inoculated with 35000HP and at 6 of 15 sites inoculated with 35000HP-SMS2 ( P = 0.14). 35000HP and 35000HP-SMS2 were recovered at similar rates from daily surface cultures and semiquantitative cultures. The data suggest that expression of MOMP is not required for pustule formation by H. ducreyi in the human model of infection.

Haemophilus ducreyi expresses a conserved hemoglobin-binding outer-membrane protein (HgbA). To test the role of HgbA in pathogenesis, we infected 9 adults with isolate 35000 and its isogenic hgbA-inactivated mutant (FX504) on their upper arms in a double-blinded, escalating dose-response study. Papules developed at similar rates at sites inoculated with the mutant or parent. The pustule-formation rate was 55% (95% confidence interval [CI], 30. 8%-78.5%) at parent sites and 0 (95% CI, 0-10.5%) at mutant sites (P

Neurofibromatosis type 1 (NF1) is a common autosomal-dominant disorder characterized by cutaneous neurofibromas infiltrated with large numbers of mast cells, melanocyte hyperplasia, and a predisposition to develop malignant neoplasms. NF1 encodes a GTPase activating protein (GAP) for Ras. Consistent with Knudson's “two hit” model of tumor suppressor genes, leukemias and malignant solid tumors in NF1 patients frequently demonstrate somatic loss of the normal NF1 allele. However, the phenotypic and biochemical consequences of heterozygous inactivation of Nf1 are largely unknown. Recently neurofibromin, the protein encoded by NF1, was shown to negatively regulate Ras activity in Nf1−/− murine myeloid hematopoietic cells in vitro through the c-kit receptor tyrosine kinase (dominant white spotting, W). Since the W and Nf1 locus appear to function along a common developmental pathway, we generated mice with mutations at both loci to examine potential interactions in vivo. Here, we show that haploinsufficiency at Nf1 perturbs cell fates in mast cells in vivo, and partially rescues coat color and mast cell defects in W41 mice. Haploinsufficiency at Nf1 also increased mast cell proliferation, survival, and colony formation in response to Steel factor, the ligand for c-kit. Furthermore, haploinsufficiency was associated with enhanced Ras–mitogen-activated protein kinase activity, a major downstream effector of Ras, via wild-type and mutant (W41) c-kit receptors. These observations identify a novel interaction between c-kit and neurofibromin in vivo, and offer experimental evidence that haploinsufficiency of Nf1 alters both cellular and biochemical phenotypes in two cell lineages that are affected in individuals with NF1. Collectively, these data support the emerging concept that heterozygous inactivation of tumor suppressor genes may have profound biological effects in multiple cell types.

At a time when more histotechnicians and histotechnologists are needed in the workforce, histology training programs are necessary. Students come to the programs with varied backgrounds, educational achievements, work experiences, and a wide range of ages. Involvement in any program that trains and graduates new histotechs is a truly rewarding experience. (The J Histotechnol 32(2):80–82, 2009)Submitted March 6, 2009; accepted with revisions April 2, 2009

A combinatorial mutagenesis strategy was used to create a collection of nearly 500 variants of human interleukin 3 (IL-3), each with four to nine amino acid substitutions clustered within four linear, nonoverlapping regions of the polypeptide. The variants were secreted into the periplasm of Escherichia coli and supernatants were assayed for IL-3 receptor-dependent cell proliferation activity. Sixteen percent of the variants, containing "region-restricted" substitutions, retained substantial proliferative activity through two rounds of screening. A subset of these was combined to yield variants with substitutions distributed through approximately half of the polypeptide. With one exception, "half-substituted" variants exhibited proliferative activity within 3.5-fold of native IL-3. A subset of the "half-substituted" variants was combined to yield "fully substituted" IL-3 variants having 27 or more substitutions. The combination of the substitutions resulted in a set of polypeptides, some of which exhibit increased proliferative activity relative to native IL-3. The elevated hematopoietic potency was confirmed in a methylcellulose colony-forming unit assay using freshly isolated human bone marrow cells. A subset of the multiply substituted proteins exhibited only a modest increase in inflammatory mediator (leukotriene C4) release. The molecules also exhibited 40- to 100-fold greater affinity for the alpha subunit of the IL-3 receptor and demonstrated a 10-fold faster association rate with the alpha-receptor subunit. The multiply substituted IL-3 variants described in this study provide a unique collection of molecules from which candidates for clinical evaluation may be defined and selected.

Background: Dermatologists have expertise in the clinical diagnosis of benign melanocytic nevi. However, there are no data to confirm the accuracy of diagnosis. Differences in the diagnostic accuracy between dermatologists and nondermatologists with regard to cutaneous tumors has been infrequently studied. Objective: We examined the rate of malignant tumors occurring in lesions submitted for routine microscopic examination that were clinically diagnosed as benign melanocytic nevi. Methods: We conducted a study at a regional, non-hospital-based dermatopathology laboratory using specimens submitted by physicians of various specialties who were practicing in a 5-state Midwest region of the United States. The preoperative and postoperative diagnoses were examined on the basis of information provided by the clinician and of the subsequent histopathologic diagnosis. A total of 7734 cutaneous pathology reports were reviewed. Specimens submitted with a preoperative clinical diagnosis of mole or nevus, with or without a modifier, were examined and compared with postoperative microscopic diagnoses. Results: Of 1946 specimens clinically diagnosed and submitted as benign nevi, 45 (2.3%) were histologically diagnosed as malignant tumors. This included 12 melanomas, 30 basal cell carcinomas, and 3 squamous cell carcinomas. For specimens submitted by dermatologists, the rate of malignant tumors increased when clinical information suggested findings beyond the classic benign clinical presentation with the addition of modifiers such as irritated or atypical , or if a malignancy was considered in the differential diagnosis (trend for increasing clinical suspicion: P = .00002). Fewer dermatologists than nondermatologists mistook a malignant tumor for a benign nevus (1.3% vs 3.8%, P = .003). Conclusion: Our data document that 2.3% of clinically diagnosed benign nevi were microscopically diagnosed as malignant tumors. Whether this malignancy rate in clinically diagnosed, benign, melanocytic nevi is above or below the threshold to establish a policy for submission for histopathologic examination remains to be determined as a collective societal and medical professional responsibility. (J Am Acad Dermatol 1999;40:567-71.)

The sequence of granulocyte colony-stimulating factor (G-CSF) has been circularly permuted by introducing new chain termini into interhelical loops and by constraining the N- and C-terminal helices, either by direct linkage of the termini (L0) or by substitution of the amino-terminal 10-residue segment with a seven-residue linker composed of glycines and serines (L1). All the circularly permuted G-CSFs (cpG-CSFs) were able to fold into biologically active structures that could recognize the G-CSF receptor. CD and NMR spectroscopy demonstrated that all of the cpG-CSFs adopted a fold similar to that of the native molecule, except for one [cpG-CSF(L1)[142/141]] which has the new termini at the end of loop 34 with the shorter L1 linker. All of the cpG-CSFs underwent cooperative unfolding by urea, and a systematically lower free energy change (DeltaGurea) was observed for molecules with the shorter L1 linker than for those molecules in which the original termini were directly linked (the L0 linker). The thermodynamic stability of the cpG-CSFs toward urea was found to correlate with their relative ability to stimulate proliferation of G-CSF responsive cells. Taken together, these results indicate that the G-CSF sequence is robust in its ability to undergo linear rearrangement and adopt a biologically active conformation. The choice of linker, with its effect on stability, seems to be important for realizing the full biological activity of the three-dimensional structure. The breakpoint and linker together are the ultimate determinants of the structural and biological profiles of these circularly permuted cytokines. In the following paper [McWherter, C. A., et al. (1999) Biochemistry 38, 4564-4571], McWherter and co-workers have used circularly permuted G-CSF sequences to engineer chimeric dual IL-3 and G-CSF receptor agonists in which the relative spatial orientation of the receptor agonist domains is varied. Interpreting the differences in activity for the chimeric molecules in terms of the connectivity between domains depends critically on the results reported here for the isolated cpG-CSF domains.

Previous work in 3 subjects infected for 2 weeks indicated that experimental infection with Haemophilus ducreyi recruits CD4 cells to the skin at the pustular stage of disease. In order to describe the kinetics of the host response, 23 subjects were infected at 2 sites with a standardized dose of H. ducreyi. Subjects were biopsied 1 or 4 days after inoculation or when they developed a painful pustular lesion (days 7-14). Papules and pustules contained a predominant T cell infiltrate that consisted of CD45RO and CD4 cells of the alpha beta lineage. Both papules and pustules contained mixed or T helper 1 type cytokine mRNA and interleukin-8 and tumor necrosis factor-alpha mRNA. Although the subjects had no history of chancroid, their immune responses resembled delayed-type hypersensitivity reactions that occurred within 24 h of inoculation and persisted throughout the course of experimental infection.

Accumulating evidence suggests an important role for the lipid mediator, platelet-activating factor (PAF), in cutaneous inflammation. In these studies the antiinflammatory effects of the potent and selective lipophilic PAF receptor antagonist A-85783 topically applied to the ventral ears of male Wistar rats were assessed. Intradermal injections of PAF into rat ears resulted in cutaneous inflammation as assessed by both ear thickness measurements and histological evaluation. Pretreatment of the ears with A-85783 resulted in an inhibition of subsequent PAF-induced inflammation. A-85783 treatment also inhibited phorbol myristic acetate-induced cutaneous inflammation, suggesting that the PAF receptor is involved in the etiology of this experimental dermatitis. These findings demonstrate that epicutaneous A-85783 is an appropriate tool to study the role of the PAF receptor in cutaneous inflammation, and suggest the possible clinical utility of this new class of antiinflammatory agents.

The balloon-cell nevus was first described over 100 years ago. Since then, balloon-cell changes of melanocytes have been noted in numerous tumors, including melanoma, blue nevus, and Spitz nevus. Whether these changes reflect cellular deterioration or proliferative changes is a matter of debate. We report a case in which balloon-cell changes were found within proliferative nodules occurring in a large congenital melanocytic nevus.

The prevalence of infections in the immunocompromised host is increasing. The oral cavity is a primary or sole site in many cases. It is important for the practicing clinician to recognize the more common infections in this growing patient population. Oral examinations are an essential component of all physical examinations, especially when immunosuppression is known or suspected. We recommend that all patients starting immunosuppressive therapy receive a comprehensive oral examination before the institution of such therapy to eliminate potential sources of oral and odontogenic infections. The examination should include full mouth dental radiographs and a complete soft-tissue examination. Timely, accurate diagnoses may have important implications with regard to management, prognosis, cost, morbidity, and mortality.

Interleukin-3 (IL-3) is a cytokine that stimulates the proliferation and differentiation of hematopoietic cells. The hyperactive hIL-3 variant SC-55494 was shown to have at least two major conformations by high-resolution NMR spectroscopy. Mutants of SC-55494 were constructed in which alanine was substituted for proline in order to test the hypothesis that proline cis-trans isomerization is the source of the observed conformational heterogeneity, as well as to evaluate the effect of prolyl peptide bond configuration on biological activity. NMR spectra of four single proline-to-alamine mutants (P30A, P31A, P33A, and P37A) retain doubled resonances, while spectra of the double mutant P30A/P31A and the quadruple mutant P30A/P31A/P33A/ P37A are substantially free of heterogeneity. These observations suggest that the two major conformations in SC-55494 correspond to cis and trans isomers of either or both of the R29-P30 and P30-P31 peptide bonds. All six mutants had somewhat lower cell proliferative activity than SC-55494, with relative activities ranging from 40 to 80%. The P37A mutant has a binding affinity to the low-affinity IL-3 receptor alpha-subunit statistically equivalent to SC-55494, while P30A, P31A, and P33A each had about two-fold decreases, and P30A/P31A and P30A/P31A/P33A/P37A had four-fold decreases. These findings suggest an important role for the cis configuration of either or both of the R29-P30 and P30-P31 peptide bonds in IL-3 for optimal interaction with the receptor alpha-subunit.