Is the 2003 ISSVD terminology and classification of vulvodynia up-to-date? A neurobiological perspective.

This paper aims to determine if the 2003 International Society for the Study of Vulvovaginal Disease (ISSVD) terminology and classification of vulval pain is up-to-date, according to a current and widely accepted neurobiological pain classification, which divides pain into nociceptive, inflammatory and pathological pain with the latter subdivided into neuropathic and dysfunctional pain. Nociceptive pain is protective, adaptive, high-threshold pain provoked by noxious stimuli. Inflammatory pain is protective, adaptive, low-threshold pain associated with peripheral tissue damage and inflammation. Pathological pain is non-protective, maladaptive, low-threshold pain caused by structural damage to the nervous system (neuropathic pain) or by its abnormal function (dysfunctional pain). The 2003 ISSVD vulval pain classification should be revised in terms of current neurobiological pain information. Inflammatory vulval pain occurs as a result of specific infectious, inflammatory and neoplastic disorders. Neuropathic vulval pain arises following a specific neurological disorder, responsible for structural damage to the nervous system. Vulvodynia is dysfunctional vulval pain, caused by abnormal function of the nervous system itself.

Provoked vestibulodynia: inflammatory, neuropathic or dysfunctional pain? A neurobiological perspective.

This paper aims to clarify the nature of the pain in provoked vestibulodynia (PV). It reviews published data about the nature of the pain in PV, employing a recent pain classification, which divides pain from a neurobiological perspective, into nociceptive, inflammatory and pathological pain, with the latter subdivided into neuropathic and dysfunctional pain. Nociceptive pain is high-threshold pain provoked by noxious stimuli; inflammatory pain is adaptive, low-threshold pain associated with peripheral tissue inflammation; pathological pain is maladaptive, low-threshold pain caused by structural damage to the nervous system (neuropathic) or by its abnormal function (dysfunctional). Most of the published data show that in PV, there is no active peripheral tissue inflammation. Similarly, no neural damage has been demonstrated. It is reasonable to consider PV as dysfunctional pain induced by exposure to acute physical or psychological precipitating events in the presence of an individual predisposition to produce or maintain abnormal central sensitisation.

The effect of somatic cell count data adjustment and interpretation, as outlined in European Union legislation, on herd eligibility to supply raw milk for processing of dairy products.

Somatic cell count (SCC) limits are a key component of national and international regulation for milk quality. As yet, very limited work has been published on SCC regulatory standards, including on the effect of different approaches to SCC data adjustment and interpretation. This study examines the effect of SCC data adjustment and interpretation, as outlined in current European Union (EU) legislation, on herd eligibility to supply raw milk for processing of dairy products for human consumption, using Irish data for illustration. The study used Irish milk-recording data as a proxy for bulk tank SCC (BTSCC) data, to calculate an unadjusted monthly SCC value for each herd during each month of participation. Subsequently, 4 data adjustments were applied, as outlined in EU and national legislation: seasonal adjustment; 3-mo rolling geometric average, without accounting for a break in the supply; 3-mo rolling geometric average, after accounting for a break in the supply; and seasonal adjustment and 3-mo rolling geometric average combined, after accounting for a break in the supply. Analyses were conducted to examine the effect, during the period from 2004 to 2010, of data adjustment on the percentage of herds with herd SCC >400,000 cells/mL. In all, 4 interpretation scenarios, incorporating different data adjustment combinations, were used to estimate herd eligibility (compliant, under warning, or suspended, as defined by legislation) to supply raw milk for processing. The 4 methods of data adjustment each led to a sizable reduction (6.7, 5.0, 5.3, and 11.1 percentage points, respectively, compared with the unadjusted data) in the percentage of herds exceeding a herd SCC of 400,000 cells/mL. Herd eligibility varied by interpretation scenarios, in particular those incorporating seasonal adjustment. The study provides new perspectives on the effect of data adjustment on herd SCC and of interpretation scenarios on herd eligibility. The results provide an illustrative, rather than definitive, picture of this effect, as national authorities use BTSCC data when determining herd eligibility, whereas this study was conducted using milk-recording data as a proxy. Some aspects of the primary EU legislation are unclear, which may lead to differences in interpretation and application. The potential impact of data adjustment and milk purchaser pricing on farm-level mastitis control in Ireland is considered.

An activated protein kinase C alpha gives a differentiation signal for hematopoietic progenitor cells and mimicks macrophage colony-stimulating factor-stimulated signaling events.

Highly enriched, bipotent, hematopoietic granulocyte macrophage colony-forming cells (GM-CFC) require cytokines for their survival, proliferation, and development. GM-CFC will form neutrophils in the presence of the cytokines stem cell factor and granulocyte colony-stimulating factor, whereas macrophage colony-stimulating factor leads to macrophage formation. Previously, we have shown that the commitment to the macrophage lineage is associated with lipid hydrolysis and translocation of protein kinase C alpha (PKCalpha) to the nucleus. Here we have transfected freshly prepared GM-CFC with a constitutively activated form of PKCalpha, namely PKAC, in which the regulatory domain has been truncated. Greater than 95% of the transfected cells showed over a twofold increase in PKCalpha expression with the protein being located primarily within the nucleus. The expression of PKAC caused macrophage development even in the presence of stimuli that normally promote only neutrophilic development. Thus, M-CSF-stimulated translocation of PKCalpha to the nucleus is a signal associated with macrophage development in primary mammalian hematopoietic progenitor cells, and this signal can be mimicked by ectopic PKAC, which is also expressed in the nucleus.

Cytokine-mediated protein kinase C activation is a signal for lineage determination in bipotential granulocyte macrophage colony-forming cells.

Granulocyte macrophage colony-forming cells (GM-CFC) have the potential to develop into either macrophages and/or neutrophils. With a highly enriched population of these cells we have found that although GM-CFC are equally responsive to macrophage colony stimulating factor (M-CSF) and stem cell factor (SCF) in terms of DNA synthesis, M-CSF stimulated the development of colonies containing macrophages in soft gel assays, while SCF promoted neutrophilic colony formation. When SCF and M-CSF were combined, mainly macrophage development was stimulated both in soft agar colony-forming assays and liquid cultures. An analysis of some potential signaling mechanisms associated with cytokine-mediated developmental decisions in GM-CFC revealed that M-CSF, but not SCF, was able to chronically stimulate phosphatidylcholine breakdown and diacylglycerol production, indicating that protein kinase C (PKC) may be involved in the action of M-CSF. Furthermore, M-CSF, but not SCF, can increase the levels of PKC alpha (PKC alpha) expression and stimulate the translocation of PKC alpha to the nucleus. When the PKC inhibitor, calphostin C, was added to GM-CFC cultured in M-CSF then predominantly neutrophils were produced, conversely PKC activators added with SCF stimulated macrophage development. The data indicate a role for PKC in M-CSF-stimulated macrophage development from GM-CFC.

The effects of exposure to an acute naturalistic stressor on working memory, state anxiety and salivary cortisol concentrations.

Exposure to an acute naturalistic stressor induces both psychological and physiological changes in humans. The two studies reported here explored the impact of exposure to an acute naturalistic stressor on state anxiety, working memory and HPA axis activation (salivary cortisol). In both experiments, ten healthy male participants were exposed to an acute naturalistic stressor, helicopter underwater evacuation training (HUET), and their physiological and behavioural responses before (first study) and after (second study) the stressor were compared to ten non-stressed controls. The results of both experiments showed that working memory performance was preserved during anticipation of an acute stressor, but impairments were observed immediately after stress exposure. Participants reported significantly higher state anxiety levels during anticipation and following stress exposure, whereas significant elevations in cortisol levels were only observed 25 min post exposure to stress, but not before or immediately after stress exposure. The results of both experiments demonstrated a dissociation between behavioural and biochemical measures and provided evidence for a dissociation of the effects of stress on cognitive and physiological measures depending on the time of testing, with cognitive impairments most evident following stress exposure.

Differential gene expression in the striatum of mice with very low expression of the vesicular monoamine transporter type 2 gene.

The vesicular monoamine transporter type 2 (VMAT2) packages pre-synaptic monoamines into vesicles. Previously, we generated mice hypomorphic for the VMAT2 gene (Slc18a2), which results in a approximately 95% reduction in VMAT2 protein, disrupted vesicular storage, severe depletion of striatal dopamine and mice with moderate motor behaviour deficits. Dopamine released from mid-brain dopamine neurons acts on post-synaptic type 1 (D1) and 2 (D2) receptors located on striatal medium spiny neurons to initiate a signalling cascade that leads to altered transcription factor activity, gene expression and neuronal activity. We investigated striatal gene expression changes in VMAT2hypo mice by quantitative real-time PCR and in situ hybridisation. Despite unaltered expression of D1 and D2 dopamine receptors, there were dramatic alterations in striatal mRNAs encoding the neuropeptides substance P, dynorphin, enkephalin and cholecystokinin. The promoters of these genes are regulated by a combination of transcription factors that includes cAMP responsive element binding protein-1 (CREB) and c-Fos. Indeed, the changes in peptide mRNAs were associated with elevated expression of Creb1 and c-Fos. These data indicate that striatal dopamine depletion, as a consequence of deficient vesicular storage in this mouse, triggers a complex program of gene expression, consistent with this mouse being an excellent model of Parkinson's disease.

Activation of the Abelson tyrosine kinase activity is associated with suppression of apoptosis in hemopoietic cells.

A chromosomal translocation uniquely associated with chronic myeloid leukemia leads to the formation of a chimeric gene, bcr-abl, on the Philadelphia chromosome. The BRC-ABL protein displays an uncontrolled tyrosine kinase activity similar to that seen with the transforming oncogene of the Abelson murine leukemia (ABL) virus (v-abl). An interleukin 3 dependent cell line, IC.DP, has been transfected with a gene encoding a temperature sensitive v-ABL. In the absence of interleukin 3 at the restrictive temperature for ABL tyrosine kinase activity IC.DP cells died via apoptosis. At the permissive temperature ABL tyrosine kinase activity promoted IC.DP cell survival but not proliferation. ABL therefore can specifically suppress apoptosis.

p210 Bcr-Abl expression in a primitive multipotent haematopoietic cell line models the development of chronic myeloid leukaemia.

Chronic myeloid leukaemia (CML) is a clonal disorder of the pluripotent haemopoietic stem cell, the hallmark of which is the constitutively activated Bcr-Abl protein tyrosine kinase. During the initial chronic phase of CML the primitive multipotent leukaemic progenitor cells remain growth factor dependent and are capable of producing terminally differentiated cells. Although the available evidence suggests that Bcr-Abl directly affects signalling pathways involved in controlling the development of primitive haemopoietic progenitors the identification of the specific biological consequences of Bcr-Abl activity in these progenitors has been hampered by the lack of suitable systems modelling CML. By transfecting the multipotent haemopoietic cell line FDCP-Mix with a temperature sensitive mutant of Bcr-Abl we have developed the first working model that mirrors the chronic phase of CML. FDCP-Mix cells expressing Bcr-Abl tyrosine kinase activity remain growth factor dependent and retain their ability to differentiate. Normal neutrophilic cells are formed in response to G-CSF and GM-CSF. In addition, the transfected FDCP-Mix cells grown at the permissive temperature for Bcr-Abl tyrosine kinase activity display enhanced survival and proliferation in low concentrations of growth factor. These findings are consistent with the initial subtle changes seen in CML progenitor cells during the chronic phase and confirm that Bcr-Abl effects are context specific, i.e. they depend on the origin and developmental potential of the transfected cells. This questions the significance of studies in non-haemopoietic and differentiation blocked haemopoietic cells.

Bcr-Abl protein tyrosine kinase activity induces a loss of p53 protein that mediates a delay in myeloid differentiation.

Chronic myeloid leukaemia is a haemopoietic stem cell disorder, the hallmark of which is the expression of the Bcr-Abl Protein Tyrosine Kinase (PTK). We have previously reported that activation of a temperature sensitive Bcr-Abl PTK in the multipotent haemopoietic cell line FDCP-Mix for short periods resulted in subtle changes including, a transient suppression of apoptosis and no inhibition of differentiation. In contrast, activation of the Bcr-Abl PTK for 12 weeks results in cells that display a delay in differentiation at the early granulocyte stage. Flow cytometric analysis also indicates that the expression of cell surface differentiation markers and nuclear morphology are uncoupled. Furthermore, a significant number of the mature neutrophils display abnormal morphological features. Prolonged exposure to Bcr-Abl PTK results in interleukin-3 independent growth and decreased p53 protein levels. FDCP-Mix cells expressing a dominant negative p53 and p53null FDCP-Mix cells demonstrate that the reduction in p53 is causally related to the delay in development. Returning the cells to the restrictive temperature restores the p53 protein levels, the growth factor dependence and largely relieves the effects on development. We conclude that prolonged Bcr-Abl PTK activity within multipotent cells results in a reduction of p53 that drives a delayed and abnormal differentiation.

Biological consequences of p160v-abl protein tyrosine kinase activity in a primitive, multipotent haemopoietic cell line.

A temperature sensitive abl protein tyrosine kinase gene was transferred into a multipotent haemopoietic stem cell line, and the primary biological effects of expression of the gene were examined at the permissive and non-permissive temperatures. Unlike previous studies in factor-dependent cell lines, we found that expression of the functional abl protein tyrosine kinase did not lead to growth autonomy. Furthermore, the cells were still able to undergo terminal myeloid differentiation. However, expression of the functional gene did lead to a delay in maturation with a concomitant increase in cell production, had a modest effect in terms of delayed apoptosis particularly when the cells were maintained at a high cell density, and slightly increased the response to sub-optimal concentrations of IL-3. In many respects, therefore, the effects of abl protein tyrosine kinase in these cells mimics the effect of bcr/abl in primary haemopoietic cells where growth factor independence and an aberrant differentiation profile are relatively late events in clonal evolution and are not intermediate consequences of activation of the abl gene.

Comparison and reproducibility of visual echocardiographic and quantitative radionuclide left ventricular ejection fractions.

Left ventricular (LV) ejection fraction (EF) is commonly assessed by equilibrium radionuclide angiography and echocardiography. These methods are presumed to be interchangeable for this purpose. This study (1) compares quantification of LVEF by equilibrium radionuclide angiography with visual estimation of LVEF by echocardiography, (2) determines the reproducibility of both methods, and (3) evaluates whether differences in determinations of LVEF are of clinical relevance. Seventy-three clinically stable patients had both equilibrium radionuclide angiography and echocardiography performed within a 4-day period. LVEF by both techniques was compared after blinded analysis by 3 echocardiographers and 3 nuclear technologists. Reproducibility was assessed by blinded repeat analysis after a 1-week interval. The frequency of differences in repeat assessments of EF that the authors considered to be of potential clinical relevance (i.e., difference > or = 10% EF units) was assessed for both techniques. Correlation of LVEF determined by both methods was good (r = 0.81, SEE = 3.5) but with substantial differences in individual patients (limits of agreement, 23.6%). Intra- and inter-observer reproducibility was good for both methods, but better for radionuclide LVEF than for echocardiographic LVEF. Limits of agreement were substantially better for radionuclide LVEF than for echocardiographic LVEF (1.8% to 3.6% versus 13.4% to 17.4%, respectively). Clinically relevant differences did not occur on repeat processing of equilibrium radionuclide angiography. In contrast, potentially clinically relevant differences occurred in 8% to 26% of studies on repeat analysis of echocardiography. Thus, LVEF determined by equilibrium radionuclide angiography and echocardiography show good agreement. Both methods provide clinically valuable measurements for LV function. However, when a precisely reproducible measurement is required for patient management decisions, equilibrium radionuclide angiography is the method of choice.

The immunoarchitecture of cutaneous pseudolymphoma.

The immunoarchitecture of five cutaneous pseudolymphomas was studied by staining serial sections for T- and B-cell and dendritic reticulum cell (DRC) antigens with monoclonal antibodies, and compared with that of reactive lymph nodes and cutaneous lymphoma. In four cases compartmentalization of B and T cells was observed, analogous to findings in reactive lymph nodes. In two of these cases the immunoarchitectural features were strikingly similar to those of reactive lymph nodes. Both had distinct follicles with germinal centers, and in one distinct mantle zone formation was seen. B cells in the follicles were polyclonal, with kappa chain predominance. The germinal centers showed the expected intercellular and/or dendritic pattern of immunoglobulin heavy chain, B2, and DRC-antigen expression. T cells admixed in the germinal centers were overwhelmingly of the T-helper type. The B-cell compartments in the other two cases showed some subtle immunologic evidence of aberrance, but the weight of evidence suggested reactive/aberrant rather than malignant processes. The T-cell compartments in all four cases showed a predominance of T-helper and a minority of T-suppressor/cytotoxic cells. All contrasted with the lymphomas, which showed B-cell monoclonality, markedly deranged T-subset proportions, or novel T-cell phenotypes. Although the main focus of this study was cases involving substantial populations of both B and T cells, preliminary observations were made in one case in which a predominance of T cells and prominent epidermotropism simulated mycosis fungoides. Quantitative ultrastructural analysis in this case suggested a reactive T-cell process. Leu-6-positive Langerhans cells were increased in the epidermis and dermis in all five cases, and in the dermis they were found almost exclusively in T-cell compartments. It is proposed that this distribution is the anatomic correlate to the known functional role of Langerhans cells in antigen processing/presentation and T-cell activation. In the cutaneous "lymph node equivalent," Langerhans cells are analogous to interdigitating reticulum cells of reactive lymph nodes in distribution and, probably, in function. The DRC found in the germinal centers in two cases were probably antigenically identical and functionally analogous to those in germinal centers of reactive lymph nodes. Immunologic phenotyping of serial cutaneous sections may aid in distinguishing reactive from neoplastic lymphoid lesions. Immunoarchitectural analysis promises to be a powerful tool for the study of lymphoproliferative disease.

Vulvar histology after neutral red photoinactivation of herpes simplex virus.

The use of photodynamic dye and light inactivation for the treatment of genital herpes simplex virus infections has been associated with the risk of potential oncogenesis. Sixteen patients treated with neutral red and fluorescent light for documented herpetic infections were studied at intervals ranging from 9 to 52 months following treatment. Four patients treated with other modalities were included in the study. Biopsies of the treated areas were obtained, and 3925 tissue sections were examined. Mild atypical epithelial changes were focally present in most specimens regardless of therapy. Histologically identifiable premalignant change could not be demonstrated.

The growth inhibitory role and potential clinical value of macrophage inflammatory protein 1 alpha in myeloid leukaemias.

The control of primitive haemopoietic progenitor cell proliferation in vitro can be achieved with combinations of growth stimulatory cytokines. Acting in apparent opposition to these growth stimulators are growth inhibitory substances, including prostaglandins, cytokines and chemokines which bind to specific cognate cell surface receptors and promote signal transduction events that interfere with cellular proliferation. Within the bone marrow microenvironment, significant quantities of both growth inhibitors and growth promoters can be detected. The ratio of their concentrations within microenvironmental niches of the marrow may regulate primitive blood cell production. The potential exists, therefore, for the disregulation of haemopoiesis via the disruption of the balance between positive and negative regulators of haemopoietic progenitor proliferation. In one particular disease, chronic myeloid leukaemia (CML), there is a lack of response of leukaemic cells to the chemokine growth inhibitor, Macrophage Inflammatory Protein-1alpha (MIP-1alpha). The role of MIP-1alpha in regulation of haemopoiesis, the response of CML progenitor cells and other myeloid leukaemic cells to this chemokine, and the reasons for lack of response to MIP-1alpha in leukaemic cells are reviewed.

Erythema nodosum, estrogens, and pregnancy.

During the past ten years we have seen two or more episodes of erythema nodosum develop in five patients in association with either pregnancy or the ingestion of female hormones. No alternative causes of erythema nodosum were found in these patients. The occurrence of erythema nodosum in association with hormonal change suggests that female hormones can, under some circumstances, be directly responsible for development of the disease.

Quantitative electron microscopy in the diagnosis of mycosis fungoides. A simple analysis of lymphocytic nuclear convolutions.

In the present study we compared the accuracy of three different ultrastructural methods used to confirm clinically proved cases of mycosis fungoides (MF) from skin biopsy specimens. Method 1 is a qualitative method and confirmed the diagnosis of MF in only 44% of the patients with MF. Method 2 involved image analysis of lymphocytic nuclei and confirmed the diagnosis of MF in only 67% of the patients with MF. The most sensitive ultrastructural method for confirming the diagnosis of MF was a simple scoring of the number of sharply angled nuclear invaginations in 100 lymphocytes (method 3). Control biopsy specimens had many more lymphocytes (19% to 55%) with no sharply angled nuclear invaginations compared with those from patients with MF (3% to 15%). The clinical diagnosis of MF was confirmed in 100% of our patients with MF using method 3.

Urticarial vasculitis treated with colchicine.

Cutaneous vasculitis may present as an urticaria-like eruption in association with viral illness, systemic lupus erythematosus, Sjögren's syndrome, or serum sickness. Often, however, urticarial vasculitis presents as an immune complex-mediated disease without other associated connective-tissue disease or associated infection. Many forms of therapy have been tried, including antihistamines, prednisone, and immunosuppressive drugs without success in every patient. Because colchicine had been effective in some patients with non-urticarial cutaneous vasculitis, we decided to use it in two patients with urticarial vasculitis in whom other therapy had failed. Both responded dramatically, although severe hypocomplementemia persisted in one patient.

An ultrastructural morphometric and immunohistochemical analysis of cutaneous lymphomas and benign lymphocytic infiltrates of skin. Useful criteria for diagnosis.

Our combined ultrastructural, immunohistochemical, histologic, and clinical studies over the past five years have allowed us to compile diagnostic criteria useful in the evaluation of cutaneous lymphomas. As a group, mycosis fungoides (MF) patients could be distinguished from those with benign disorders of skin using ultrastructural morphometry (mean form factor and perimeter values), but with some overlap between groups. Another approach, the ultrastructural histogram method, however, clearly separated MF patients from patients with chronic dermatitis. Immunohistochemistry was useful in distinguishing cases of cutaneous peripheral T-cell lymphoma from MF cases on the basis of the occurrence of "novel phenotypes." Neoplastic T-cell infiltrates of skin can usually be distinguished from benign polyclonal T-cell infiltrates by the presence of one T-cell subset to the exclusion of others. Patients with convoluted B-cell lymphomas could also be distinguished from MF patients using ultrastructural morphometric dual parameter analysis. The diagnostic complexity of several cutaneous T-cell lymphoma cases is illustrated. We have emphasized in this study the strength of combining quantitative electron microscopy, immunohistochemistry, and histology in the diagnostic workup of cutaneous lymphomas. This integrative approach may be necessary to assure a definitive diagnosis in difficult cases.

Intralesional triamcinolone therapy for pretibial myxedema.

In a prospective study, nine patients with pretibial myxedema were treated with intralesional injections of triamcinolone acetonide. Complete resolution of the myxedematous plaques was obtained in seven of the nine patients. The other two patients failed to complete their treatment programs but did show partial resolution. For most patients the monthly injection of 8 ml or less of a solution containing 5 mg/ml of triamcinolone proved to be the most effective dosage schedule. No serious side effects were encountered. New nodules of myxedema developed in some patients after the initial completion of therapy; these nodules responded to reinjection using the same dosage schedule.