Ichthyosiform eruptions in association with primary cutaneous T-cell lymphomas.

BACKGROUND: Malignant lymphoma is occasionally complicated by ichthyosiform eruptions. OBJECTIVES: To analyse histopathologically the ichthyosiform eruptions associated with cutaneous lymphomas. METHODS: We reviewed the files of patients with malignant lymphoma seen in our dermatology department between January 2001 and May 2006 to search for patients with ichthyosiform eruptions. RESULTS: In our series, nine of 106 patients with malignant lymphomas had ichthyosiform eruptions during their clinical courses, including three (30%) of 10 patients with anaplastic large cell lymphoma (ALCL) and six (14%) of 44 patients with mycosis fungoides (MF). None of the 18 patients with cutaneous B-cell lymphoma had ichthyosiform eruptions. The three patients with ALCL had ichthyosiform eruptions histopathologically consistent with acquired ichthyosis (AI) in which packed horny layers and thin granular layers were present without lymphocytic infiltration. In contrast, four of the six patients with MF (stages Ib and IIb) had ichthyosiform eruptions with epidermotropic infiltration of atypical lymphocytes, as observed in ichthyosiform MF (IMF). Of the remaining two patients, one showed histopathological features overlapping AI and IMF, and the other had AI alone. These two patients (stages IVa and IIb) had tumours composed of CD30+ cells. Filaggrin expression was markedly diminished in both AI and IMF-like eruptions, similar to that of inherited ichthyosis vulgaris. CONCLUSIONS: Ichthyosiform eruptions are often associated with ALCL and MF and can be classified into three groups: AI associated with ALCL and MF expressing CD30, IMF, and their overlap.

Human papillomavirus genome integration in multifocal vulvar Bowen's disease and squamous cell carcinoma.

Human papillomavirus (HPV) is an important aetiological agent in cervical carcinomas and in malignant skin tumours. Integration of the HPV DNA into host genome is one of the most important risk factors for malignant transformation. We report a patient with multiple black plaques and an erythematous nodule on her vulva. On histological examination, multifocal vulvar Bowen's disease (BD) and invasive squamous cell carcinoma (SCC) were found. An amplification of papillomavirus oncogene transcripts (APOT) assay showed that two locations of BD had only episome-derived HPV16 transcripts, but the other two sites of BD and the nodule of invasive SCC had HPV16 transcripts derived from integration. Sequencing analysis revealed that the invasive SCC had its integration site at 8q24, the Myc locus. Our results suggest that the APOT assay in multiple sites of the same patient may be a valuable tool for evaluation of the clinical degree of malignancy for vulvar BD.

Leukotriene B4-activated human endothelial cells promote transendothelial neutrophil migration.

We explored the effect of leukotriene B4 (LTB4) on endothelial cells in LTB4-induced transendothelial migration (TEM) of neutrophils as an in vitro model of neutrophil extravasation. Chemotactic response of human neutrophils to LTB4 was significantly lower than that in response to N-formyl-methionyl-leucyl-phenylalanine (fMLP), whereas the extent of TEM in response to LTB4 was significantly higher than that to fMLP. The study on random migration induced by LTB4 and fMLP also showed similar results, which indicated that LTB4 might affect the human umbilical cord vein endothelial cell (HUVEC) barrier. Neutrophil TEM was induced by pretreatment of HUVEC monolayer with LTB4 but not with fMLP. Treatment of endothelial cells by ONO-4057, a LTB4 receptor antagonist, abolished the effect of LTB4 almost completely whereas neutrophils treated with ONO-4057 could transmigrate through HUVEC treated with LTB4. These findings indicated that LTB4 could induce neutrophil TEM by acting on HUVEC.

Random migration of polymorphonuclear leukocytes induced by GM-CSF involving a signal transduction pathway different from that of fMLP.

Granulocyte-macrophage colony-stimulating factor (GM-CSF) induced random migration of human polymorphonuclear leukocytes (PMNs) but not chemotaxis. Chemoattractants such as N-formyl-methionyl-leucyl-phenylalanine (fMLP), leukotriene B4 (LTB4), and interleukin-8 (IL-8) induced both random migration and chemotaxis. Other inflammatory cytokines, including granulocyte colony-stimulating factor (G-CSF), interleukin 1alpha (IL-1alpha), and tumor necrosis factor alpha (TNF-alpha), did not induce either movement. One-minute exposure of PMNs to GM-CSF was sufficient for the induction of random migration, whereas fMLP-induced random migration required continued presence of fMLP. Inhibitors of phosphatidylinositol 3-kinase (PI3-K), protein kinase C (PKC), and protein tyrosine kinase (PTK) had no effect on random migration induced by GM-CSF, whereas fMLP-induced movements were partially inhibited by PTK inhibitors but not by inhibitors of PI3-K inhibitors nor PKC inhibitors. Myosin light chain kinase inhibitors inhibited movements of PMNs induced by both GM-CSF and fMLP. These findings also imply that some aspects of the signal transduction pathway of GM-CSF leading to random migration is different from that of fMLP. Our findings suggest that cell movements are controlled through diverse signal transduction systems.

Versican upregulation in Sézary cells alters growth, motility and resistance to chemotherapy.

Sézary syndrome (SéS) represents a leukemic variant of cutaneous T-cell lymphoma, whose etiology is still unknown. To identify dyregulated genes in SéS, we performed transcriptional profiling of Sézary cells (SCs) obtained from peripheral blood of patients with SéS. We identified versican as the highest upregulated gene in SCs. VCAN is an extracellular matrix proteoglycan, which is known to interfere with different cellular processes in cancer. Versican isoform V1 was the most commonly upregulated isoform in SCs. Using a lentiviral plasmid, we overexpressed versican V1 isoform in lymphoid cell lines, which altered their growth behavior by promoting formation of smaller cell clusters and by increasing their migratory capacity towards stromal cell-derived factor 1, thus promoting skin homing. Versican V1 overexpression exerted an inhibitory effect on cell proliferation, partially by promoting activation-induced cell death. Furthermore, V1 overexpression in lymphoid cell lines increased their sensitivity to doxorubicin and gemcitabine. In conclusion, we confirm versican as one of the dysregulated genes in SéS and describe its effects on the biology of SCs. Although versican overexpression confers lymphoid cells with increased migratory capacity, it also makes them more sensitive to activation-induced cell death and some chemotherapeutics, which could be exploited further for therapeutic purposes.

Demonstration of functionally distinct human polymorphonuclear leukocyte fractions by simultaneous measurement of phagocytosis and oxygen radical generation.

Phagocytosis and oxygen radical generation by human polymorphonuclear leukocytes (PMNLs) were studied by a two-color flow cytometric analysis, where the red fluorescent product(s) of hydroethidine was used as an indicator of intracellular generation of oxygen radicals and opsonized zymosan (OZ) as an indicator of phagocytosis. Unstimulated cells formed a single population of cells without any significant fluorescence. PMNLs stimulated by OZ exhibited a high red fluorescence. Most PMNLs phagocytosed OZ and generated oxygen radicals when stimulated by zymosan particles opsonized with human AB serum at concentrations of more than 10%, whereas three distinct subpopulations (designated as R1, R2 and R3) appeared when stimulated by zymosan particles opsonized with 3% serum; R1 cells enhanced neither green nor red fluorescence, R2 cells enhanced green fluorescence but not red fluorescence, and R3 cells enhanced both green and red fluorescence. The R2 cells completely disappeared by the addition of Trypan blue. Most of the R3 cells disappeared by the addition of cytochalasin B. These findings on the three fractions were confirmed by the observation under fluorescence microscopy of cells in each fraction obtained by sorting. In conclusion, PMNLs could be separated into three functionally distinct subpopulations when stimulated by zymosan particles opsonized with a suboptimal concentration of serum.

Histamine chloramines have a persistent stimulating effect on histamine H2 receptors and gastric acid secretion.

Histamine plays an important role in the control of gastric acid secretion. Recently, chlorinated derivatives of histamine have been identified as having multiple effects on the intestinal tract. The aim of this study was to investigate the role of histamine chloramines on gastric acid secretion. We compared the effects of histamine and histamine chloramines on the histamine H2 receptors in vitro using guinea pigs and on gastric acid secretion in rats. With respect to the effects on histamine H2 receptors, histamine monochloramine showed agonist effects similar to those seen with histamine, but the agonist effects of histamine dichloramine were about half those of histamine. Unlike histamine effects, the histamine H2 receptor agonist effects of histamine monochloramine and histamine dichloramine did not disappear after repeated washout. With respect to the stimulation of gastric acid secretion in vivo, histamine monochloramine was similar to histamine, while the effect of histamine dichloramine was 42.2-52.7% of that of histamine. The recovery time to the basal secretory level after completion of stimulation by histamine chloramines was significantly prolonged compared with histamine. These results suggest that histamine chloramines, which bind strongly with histamine H2 receptors, may delay the termination of gastric acid secretion and increase the burden on the gastric and duodenal mucosa.

Effect of gastric mucosal blood flow on the pathogenesis of ammonia in Helicobacter pylori-induced gastric mucosal injury.

PURPOSE: We investigated the effects of the gastric mucosal blood flow on the pathophysiology of ammonia-induced gastric mucosal damage. DESIGN: The study was designed to show whether the damaging effect of ammonia, a product of Helicobacter pylori urease, on the gastric mucosa is increased by the decrease in gastric mucosal blood flow in rats subjected to ischemia. RESULTS: Although ammonium chloride at concentrations of 15-60 mmol/l produced no significant macroscopical lesion in normotensive rats, it caused severe macroscopic hemorrhagic gastric lesions in the stomachs of rats subjected to ischemia. Exposure of the stomach to the combination of ischemia and ammonium chloride (60-600 mmol/l, pH adjusted to 7-9 with sodium hydroxide) produced macroscopic hemorrhagic lesions. However, exposure of the mucosa to the combination of ischemia and sodium hydroxide (60-600 mmol/l, pH was reduced to 7-9 with hydrochloric acid) produced no significant lesions. CONCLUSIONS: These results indicate that the mucosal lesions observed in patients infected with H. pylori are not caused by the alkalinity of H. pylori-produced ammonia and suggest that concomitant infection with H. pylori in the ischemic stomach may lead to the development of ulceration in these patients.

Koebner phenomenon on skin graft donor site in cutaneous angiosarcoma.

An 81-year-old woman developed a necrotic plaque and a surrounding purple-red, irregularly shaped macule on her scalp. The diagnosis of angiosarcoma was confirmed histologically. A wide surgical excision was made followed by a split-thickness skin graft from her right buttock. Nine months later, she noticed a dark purple-red lesion on the donor site which grew rapidly into a large mass. Histological examination revealed irregular clefts and vascular channels lined by atypical endothelial cells. Lung metastasis and pneumothorax were also noted. The secondary tumor appeared to represent Koebner phenomenon in a patient with angiosarcoma of the scalp.

Perforator-based flap for coverage of lumbosacral defects.

Lumbosacral defects on 20 patients were covered with a perforator-based flap. Cutaneous perforators derived from the 9th and 10th intercostal arteries, the 4th lumbar artery, and multiple gluteal perforators that penetrate the gluteus maximus muscle were used as vascular pedicles. Minor complications occurred in five cases. Using this method, minimal morbidity of the donor site is expected because the gluteus maximus need not be sacrificed. Accordingly, perforator-based flaps are especially indicated for ambulatory patients, but for paraplegic patients as well. Even in the event of recurrence, another perforator-based or musculocutaneous flap can be elevated from the ipsilateral side because of the presence of multiple perforators in the lumbosacral and gluteal regions.

[Analysis of platelet-derived factors that modulate functions of polymorphonuclear leukocytes].

Interactions between platelets and polymorphonuclear leukocytes (PMN) modulate their functions and play a role in the development of pathogenesis of some disease. Platelets secret various kinds of factors that affect PMN functions. They seemed to have important role in vivo, but little has been elucidated on exact mechanism of action and physiological meaning of each factor in relation to PMN functions. We studied the effects of platelets and released substances from activated platelets on the functions of PMN. Results were as follows. 1) Platelets enhanced bactericidal activities of PMN against E.coli. 2) Platelets had effects on the generation of superoxide anion (O2-) of PMN. Their effects were quite different according to the assay condition of PMN, that is, platelets inhibited O2- generation when PMN were at rest or stimulated slightly and they enhanced O2-generation of PMN that were stimulated with optimal condition. 3) Thrombin-activated platelets and their supernatant elicited a transient elevation of [Ca2] of PMN. The activity of the supernatant decreased by treating with hexokinase that decomposed ATP. Further treatment with trypsin abolished its activity almost completely. Considering with our additional experiments, factors that induced [Ca2+] elevation of PMN were ATP, beta-thromboglobulin and some trypsin-sensitive factor(s). 4) Supernatant of thrombin-activated platelets decreased random migration and chemokinesis of PMN.

[Complete remission in acute myeloblastic leukemia (M0) after treatment with rhG-CSF].

A 57-year-old man was admitted because of fever and night sweat. The bone marrow was hypercellular with 86.4% blast cells. The diagnosis of AML (M0) was made, because the blast cells were negative for peroxidase stain and had CD13 and no lymphoid antigens in marker analysis. The patient was treated with BH-AC.TMP, BH-AC.MVP and low dose Ara-C without any hematological improvement, and even additional treatment with medium dose Ara-C resulted in 66.4% blast cells in the bone marrow. Subsequent administration of rhG-CSF (150 micrograms/day) by continuous intravenous infusion resulted in the decrease of the blast cells in the bone marrow to a level that was evaluated as complete remission. He remains in complete hematological remission at present. As shown in this case, rhG-CSF might be an effective agent for the treatment of AML, even if the mechanism of its effectiveness is unclear at present. Further clinical studies should will supply useful information to analyze the pathophysiology of AML.

[Successful treatment with donor leukocyte transfusion followed by interferon-alpha in a patient with relapsed chronic myelogenous leukemia after allogeneic bone marrow transplantation].

A 44-year-old man with CML in chronic phase was admitted for BMT from an HLA-identical sibling. Ph positive cells were undetectable at 3 and 7 months after BMT but became detectable by cytogenetic analysis of bone marrow aspirates at 12 months after BMT. He was treated with IFN-alpha (6 million units/day, 3 times a week) without apparent effect. Donor leukocyte transfusion (DLT) was performed four times between 20 months and 23 months after BMT, transfusing 3.4 x 10(8) mononuclear cells/kg. However, leukocytosis appeared and the NAP score declined at 25 months after BMT. FISH analysis revealed an increase in bcr-abl positive cells. IFN-alpha was restarted using the same schedule at 26 months after BMT. Three months after restarting IFN-alpha, the leukocyte count fell to the normal range, NAP score increased to a normal level, and bcr-abl positive cells decreased markedly. He has remained in hematological and cytogenetic remission for 20 months, and bcr-abl chimeric mRNA remained undetectable by PCR. These results suggest that CML which does not respond to DLT may be cured by subsequent IFN-alpha therapy, possibly by inducing anti-leukemia immune responses.

[A clinical study on the cases of ischemic colitis: comparison of clinical images based on time from onset of the disease to detection of bloody stool].

A study was performed on 28 cases of ischemic colitis. The patients were divided into three groups: Group A (9 cases with bloody stool detected within 2 hours after the onset of abdominal pain), Group B (7 cases with bloody stool detected in 2 to 6 hours), and Group C (12 cases with bloody stool detected after more than 6 hours). These cases were comparatively studied. Variables used were as follows: (1) age, (2) sex, (3) constipation, (4) vomiting, (5) peak value of WBC count (/microliter) after admission, (6) peak value of log CRP (microgram/dl), (7) presence of ulcerative lesion in endoscopic findings in acute stage. Using Group A B and C as classification variables, canonical discriminant analysis was performed. As a result, clear linearity was recognized in Group A-->B-->C, and the values (5), (6) and (7) were extracted as the corresponding variables. For these variables, significant difference was also noted in multivariate analysis of covariance. These results suggest that it is possible to predict the severity of the disease to some extent as represented by objective markers of inflammation by finding the time from onset of abdominal pain to detection of bloody stool.

Infectious complications and managements for surgical site infections in genital Paget's disease.

BACKGROUND: Extramammary Paget's disease is an intra-epidermal carcinoma that occurs preferentially in genital areas. Patients with genital Paget's disease (GPD) sometimes develop severe post-surgical infections because of this anatomical disadvantage. OBJECTIVE: To study perioperative micro-organisms and surgical site infection (SSI) in GPD. METHODS: We examined micro-organisms isolated from preoperative lesions, necrotic sites and infected wounds in 60 adult patients with GPD who underwent surgery at our hospital between November 1990 and December 2005. Based on the obtained microbiological data, we assessed the incidence, risk factors and treatment of SSI. RESULTS: The colonized organisms found in preoperative GPD were Enterobacteriaceae (27.6%), methicillin-sensitive Staphylococcus aureus (MSSA) (22.4%) and coagulase-negative staphylococci (CNS) (15.5%), among others. In the postoperative necrotic sites, the frequency of MSSA isolation was reduced to 9.4%, while Pseudomonas aeruginosa and methicillin-resistant S. aureus (MRSA) both increased in frequency from 3.4% and 0% upon preoperative examination to 18.8% and 9.4%, respectively. The incidence of SSI was 15%. In 7 of 9 SSIs, MRSA and/or P. aeruginosa were isolated. CONCLUSION: We have successfully identified a number of perioperative micro-organisms in GPD. The present observations may be extremely useful in choosing appropriate antimicrobial agents for use in the surgical treatment of GPD.

Products of neutrophil metabolism increase ammonia-induced gastric mucosal damage.

Recent studies have indicated that ammonia is involved in the pathophysiology of Helicobacter pylori-associated gastric mucosal damage. Helicobacter pylori-associated chronic active gastritis is characterized by an invasion of neutrophils. We investigated the interrelationship among hypochlorous acid (oxidant produced by neutrophil), ammonia (product of Helicobacter pylori urease), and monochloramine (product of ammonia and hypochlorous acid) in the development of gastric mucosal damage in rats. Gastric mucosal lesions were produced by exposure of the gastric mucosa to ammonia, urea with urease, or urea with Helicobacter pylori in rats subjected to ischemia. Pretreatment with taurine (scavenger of hypochlorous acid) or antineutrophil serum significantly attenuated gastric mucosal lesions induced by the above test agents. Ammonia-induced gastric mucosal lesions were exacerbated in the presence of hypochlorous acid with concomitant generation of monochloramine. These results suggest that the ammonia, hypochlorous acid, and monochloramine triad may be important in Helicobacter pylori-mediated gastric mucosal damage.

Gastric ammonia has a potent ulcerogenic action on the rat stomach.

BACKGROUND: The pathophysiological mechanism by which Helicobacter pylori induces mucosal injury has not been clarified. The aim of this study was to investigate the role of urea, urease, and ammonia in rat gastric mucosal lesions using an ex vivo chamber model. METHODS: Two groups of rats, normotensive rats and those subjected to ischemia, were studied. The gastric mucosa was examined histologically and macroscopically, and the transmucosal potential difference was measured. RESULTS: Instillation of urea into the stomach generated ammonia in the presence of urease. The amount of ammonia was increased depending on the concentration of urea and was closely associated with the severity of the histological lesions. The exposure of the stomach to 15-60 mmol/L ammonium hydroxide induced both a reduction in transmucosal potential difference and microscopic damage to the gastric mucosa in normotensive rats. Moreover, 15-60 mmol/L ammonium hydroxide produced severe macroscopic gastric lesions in the rats subjected to ischemia. CONCLUSIONS: These results show that ammonia is deleterious to the gastric mucosa and suggest the importance of urea, urease, and ammonia in the pathophysiology of gastric diseases in H. pylori-infected patients.

Helicobacter pylori has an ulcerogenic action in the ischemic stomach of rats.

Helicobacter pylori (H. pylori) is now accepted as an important cause of chronic active gastritis. There also seems to be an association between the colonization of H. pylori in the gastric mucosa and peptic ulceration. However, it has not demonstrated that the instillation of H. pylori into the stomach produces the ulcerative gastric lesions in animals or humans. We carried out an experiment to study whether or not H. pylori has an ulcerogenic action in the ischemic stomach of rats, using an ex vivo gastric chamber. The rat stomachs were exposed to 1 ml of H. pylori solution (200 IU of urease/ml) and 1 ml of urea (400 mg/dl) for 60 min after the creation of ischemia in the stomach (by withdrawal of 3 ml of blood). The exposure of the stomach to both H. pylori and urea resulted in severe hemorrhagic gastric mucosal lesions with a marked decrease in potential difference (PD) with a concomitant increase in ammonia concentration in rats with ischemia, whereas gastric lesions and a fall in PD were hardly observed in rats without ischemia. These results have demonstrated that H. pylori has an ulcerogenic action on the stomach subjected to mucosal ischemia.

Combined anterior thigh flaps and vascularised fibular graft for reconstruction of massive composite oromandibular defects.

Six massive, composite oromandibular defects were reconstructed using combined anterior (anterolateral and anteromedial) thigh flaps and vascularised fibular graft in bridge or chimeric fashion. Except for minor dehiscence in one case and infection in another, all flaps survived without complication. Anterior thigh flaps provide large-calibre, long vascular pedicles while derivative branches from the lateral circumflex femoral system facilitate simultaneous transplantation of multiple components. Because the pedicle of the vascularised fibular graft is of insufficient length, the lateral circumflex femoral vessels provide a remote vascular source as a flow-through vascular conduit. A combined flap using the lateral circumflex femoral system is considered to be most suitable for reconstruction of through-and-through defects of the head and neck. The authors describe the advantages of this method and the detailed anatomy of the cutaneous perforators of the anterior thigh flaps.