Detection of viral DNA within skin of healed recurrent herpes simplex infection and erythema multiforme lesions.

The polymerase chain reaction (PCR) was used to detect HSV DNA in genomic DNA extracted from skin biopsies obtained from healed skin of five patients with hyperpigmented macules following recurrent cutaneous HSV infections and from eight patients with HSV-associated erythema multiforme (EM). A 92-bp HSV-1 DNA fragment was found in all the skin biopsies from the site of recurrent HSV infection and in five of eight (62%) biopsies from the EM patients. Virus DNA was not found in tissues distant from the site of HSV recurrence or from a patient without a history of HSV infection. These findings confirm the presence of HSV in healed skin from the site of recurrent HSV disease and are consistent with the concept that HSV is involved in EM pathogenesis.

A novel gene expressed in human keratinocytes with long-term in vitro growth potential is required for cell growth.

The herpes simplex virus large subunit of ribonucleotide reductase differs from its counterparts in eukaryotic and prokaryotic cells and in other viruses in that it contains a unique domain that codes for a distinct serine-threonine protein kinase that activates the Ras/MEK/MAPK mitogenic pathway and is required for virus growth. Previous studies suggested that ribonucleotide reductase protein kinase was co-opted from a cellular gene. Cellular genes similar to ribonucleotide reductase protein kinase were not cloned, however, and their function is unknown. Here we report that a novel gene (H11) that codes for a protein similar to herpes simplex virus 2 ribonucleotide reductase protein kinase, is expressed in skin tissues, cultured keratinocytes, and the keratinocyte cell line A431. The protein is phosphorylated and it associates with the plasma membrane. H11 is expressed in keratinocytes with long-term in vitro growth potential and is coexpressed with high levels of adhesion molecules involved in signal transduction, such as beta1 integrin. Antisense oligonucleotides that inhibit H11 expression inhibit DNA synthesis and keratinocyte proliferation, suggesting that H11 expression is required for cell growth.

Lymphomatoid papulosis. Case reports and literature review.

Five new cases of lymphomatoid papulosis are reported. This entity can apparently be divided into at least two subtypes. In one group of patients, papulopustular lesions resembling those of pityriasis lichenoides develop that resolve spontaneously within a few weeks. The lesions contain an epidermotropic dermal infiltration composed predominantly of abnormal lymphocytes. The other group of patients displays larger, more persistent lesions that contain a nonepidermotropic dermal infiltration composed predominantly of histiocytes. Lymphoma eventually develops in about 10% of the patients with lymphomatoid papulosis.

Angiolymphoid hyperplasia with eosinophilia.

Angiolymphoid hyperplasia with eosinophilia is manifested by benign vascular tumors that usually occur about the head in young adults. Clinical and histologic variation of this process has resulted in nosologic confusion, and the cases in the English literature were reviewed to characterize it within racial groups. Oriental patients tend to be younger at onset, usually male, and have marked blood eosinophilia; large, but relatively asymptomatic, tumors develop in these patients. White patients tend to be older at onset and have small, friable tumors. Large tumors that are painful or pruritic tend to develop in blacks and Middle Eastern patients.

Teledermatology and in-person examinations: a comparison of patient and physician perceptions and diagnostic agreement.

OBJECTIVE: To compare physician and patient impressions and interphysician diagnostic agreement between live teledermatology and in-person examinations. DESIGN: Paired video and in-person examinations with different dermatologists. SETTING: An urban Veterans Affairs dermatology clinic. PATIENTS: One hundred thirty-nine patients. MAIN OUTCOME MEASURES: Satisfaction questionnaires and interphysician diagnostic agreement. RESULTS: Patient and physician satisfaction was high. Agreement between video and in-person diagnoses was 80%. CONCLUSIONS: Physicians and patients were satisfied with teledermatology examinations. Diagnostic agreement between in-person and video dermatologists was high.

Partial purification of Chironex fleckeri (sea wasp) venom by immunochromatography with antivenom.

Chironex fleckeri crude venom was partially purified using immobilized commercially available ovoid antivenom. The antibody preparation reacted with lethal, hemolytic, dermonecrotic and mouse writhing (pain) factors in the crude venom. The lethal activity was purified five fold, while the specific eluate contained lower quantities of hemolytic, dermonecrotic and mouse writhing activities than did the crude venom.

Pharmacological effects of various venoms on cutaneous capillary leakage.

Studies to counteract the cutaneous vasopermeability actions of a wasp (Vespa orientalis), an anemone (Bolocera tuediae) and three jellyfish (Chironex fleckeri, Chrysaora quinquecirrha and Physalia physalis) venoms were conducted by using various pharmacological antagonists. Piripost (a leukotriene inhibitor) reduced vasopermeability if administered 5 min prior to challenge with the jellyfish venoms. Methysergide counteracted the vasopermeability of three of four coelenterate venoms, whereas indomethacin was effective against capillary leakage induced by Chironex venom. These studies indicate that anti-dermonecrotic therapy against various venoms will have to be species-specific.

Techniques applicable for purifying Chironex fleckeri (box-jellyfish) venom.

A survey of several techniques to isolate a purified lethal factor from the tentacles of Chironex fleckeri was completed. Heterologous band patterns were obtained from specific eluates after gel filtration, ion exchange, immunoaffinity and hydrophobic chromatography. SDS-PAGE revealed a dense band at 24,000 mol. wt in many of these fractions. Isoelectric focusing of the crude venom resulted in considerable loss of activity but indicated significant purification in the fractions having a pI of 5.2-6.8. These fractions were also immunologically active against sera from a convalescing post-evenomation patient. The primary difficulties encountered in jellyfish venom purification are the lack of stability and the tendency of the active toxins to adhere to each other and to various support matrices.

Prolonged blurred vision following coelenterate envenomation.

The case history of a healthy woman who developed persistent (8 days) blurred vision within 3 hr of a jellyfish (Linuche unguiculata) sting on her knee is reported. Persistent sequelae following envenomation by marine animals can occur but are unusual.

A fatal jellyfish envenomation by the Portuguese man-o'war.

The documented case of a human fatality resulting from envenomation by Physalia physalis is presented. Although this case is only the second scientifically recorded fatality, several similar cases have been suspected.

Venomous pelagic coelenterates: chemistry, toxicology, immunology and treatment of their stings.

Ten years have elapsed since our last review article on the toxicology of venomous pelagic coelenterates was published (Burnett and Calton, 1977). Investigation on important medusae and the chemistry of their nematocyst venoms have been expanding. The venomous jellyfish discussed here include the Portuguese man-o'war, (Physalia physalis), the sea nettle (Chrysaora quinquecirrha), the box jellyfish (Chironex fleckeri and/or Chiropsalmus quadrigatus), the cabbage head jellyfish (Stomolophus meleagris), the lion's mane jellyfish (Cyanea capillata), the Irukandji jellyfish (Carukia barnesi), the Moreton Bay Carybdeid medusa (Morbakka), and the mauve blubber (Pelagia noctiluca).

Recurrent eruption following a solitary envenomation by the cnidarian Stomolophous meleagris.

The case history of a patient who developed a linear erythematous eruption on the ankle after envenomation by Stomolophous meleagris is described. This eruption subsided within 20 hr and recurred without subsequent envenomation 7 days later. The recurrent eruption was similar in morphology to the original lesion, but was accompanied by stiffness and edema. Examination of biopsied skin from the recurrent eruption showed a perivascular mononuclear cell dermal infiltrate. No abnormal deposits of immune globulins or complement were found in this specimen. The patient's serum contained antibodies to Stomolophous antigen and two other jellyfish. Two screening tests of lymphocyte function were negative upon exposure to Stomolophous antigen. This was the first reported case of recurrent local eruption to a single human envenomation by this coelenterate, which usually is regarded as relatively harmless to man.

Toxinological studies on the nematocyst venom of Chrysaora achlyos.

Nematocyst venoms from both oral arms and lappets of Chrysaora achlyos were prepared and found to have factors producing mouse lethality, hemolysis and hepatocyte toxicity. These venoms had less potency than those of Chrysaora quinquecirrha a phylogenetic, congeneric cousin. Envenomated bathers had significant species-specific anti venom IgG and also cross-reacting antibody to Chrysaora quinquecirrha nematocyst venoms. There were similarities and contrasts in the capillary electropherograms and sodium dodecyl sulfate (SDS) gels between C. achlyos nematocyst venoms and those of their C. quinquecirrha counterparts.

Characteristics of hyaluronidase and hemolytic activity in fishing tentacle nematocyst venom of Chrysaora quinquecirrha.

The enzyme, hyaluronidase, detected in crude venom was active over a pH range of 4-9 and was stable to at least a 72 hr storage at 4 degrees C. Preparative electrofocusing indicated a pI value for this enzyme between 9.5 and 10.2. Hyaluronidase partially purified by gel-filtration chromatography was evaluated as a spreading factor for dermonecrosis provoked by the nematocyst venom on depilated rats. The spread of dermonecrosis increased regardless of the presence or absence of hyaluronidase. Hemolytic activity of crude venom was assayed on erythrocyte suspensions from various sources. Pig and rat erythrocytes were the most sensitive to the hemolysin of the erythrocytes tested. The pH optima for the hemolysin was 8.3. Trypsinized rat erythrocytes were more susceptible to hemolysis induced by venom than non-trypsinized cells. The monovalent and divalent cations KCl, BaCl2, CaCl2, and MgCl2 were inhibitory to hemolytic activity induced by crude venom. The hemolysin partially purified by gel-filtration chromatography tested for stability after overnight storage at 4 degrees C and -70 degrees C indicated a 50% and 75% loss of activity, respectively, in comparison to the hemolytic activity recovered directly after gel-filtration chromatography.

Sea nettle (Chrysaora quinquecirrha) lethal factor: purification by recycling on m-aminophenyl boronic acid acrylic beads.

Experiments were performed to define the best isolation method for isolating Chrysaora fishing tentacle nematocyst organelles in order to minimize non-nematocyst contaminating proteins and proteases and stabilize crude nematocyst venom lethal activity. Techniques employed to disrupt the tentacles included autolysis, homogenization, or digestion using either trypsin or collagenase. Sephacryl-200 gel-filtration chromatography separated two lethal fractions. An immobilized serine protease inhibitor column, m-aminophenyl boronic acid acrylic beads, which reversibly bound one of the two lethal factors, was used in the second and third purification steps. By this means, a 105,000 mol wt. protein was purified, as judged by silver stained SDS-polyacrylamide gels. Lethal activity was inhibited by exposure to the serine protease inhibitor, L-1 chloro 3[4-tosylamido]-7-amino-2-heptanone-HCl, after purification. Although this lethal factor has some characteristics of a serine protease, it is not proteolytically active.

Lymphokine activity in coelenterate envenomation.

Four patients were tested to determine whether lymphokine activity could be produced by their peripheral blood leukocytes exposed to jellyfish antigens. One patient was a false positive reactor, one had a prolonged lymphokine response three years after a persistent eruption induced by the venom, and one had lymphokine activity associated only with the cutaneous eruption. These results support the hypothesis that T cell function may be important in the pathogenesis of cutaneous lesions following coelenterate envenomation.

Human immunosuppression induced by sea nettle (Chrysaora quinquecirrha) venom.

Large intradermal injections of crude sea nettle (Chrysaora quinquecirrha) venom in normal saline produced immunosuppression in a healthy human adult male. This response persisted several days and was homologous against that coelenterate antigen but also heterologous against antigens contained within vaccinia and herpes simplex viruses and tetanus bacillus. This down-regulation of immunity was probably mediated by cells with functional properties of suppressor cells and could be reversed by indomethacin suggesting a role for prostaglandins. This result suggests the possibility that naturally occurring environmental agents, other than sunlight, may influence the human immune response.

Partial purification of box jellyfish (Chironex fleckeri) nematocyst venom isolated at the beachside.

Chironex fleckeri, the northern Australian box jellyfish produces one of, if not, the most potent animal venoms. Study of the venom has been hampered by the limits of the animals' range and the venom's thermolability. Using retained lethality and native polyacrylamide gel electrophoresis (NPAGE), we show that lyophilization of autolysis isolated nematocysts is an effective method of transporting the venom. In addition, Sephadex G-200 chromatography, spin concentration, and NPAGE fail to demonstrate the presence of a 600 kDa protein to which the bulk of the lethal activity has been ascribed. Sodium dodecyl sulfate capillary electrophoresis of crude venom yields several protein bands with a molecular weight range of 30-200 kDa. Freeze-thaw studies show a loss of activity and NPAGE bands after two freeze thaw cycles.

Immunity to jellyfish venoms: suppression of venom-induced immune responses in ultraviolet B-irradiated mice.

Lymph node and spleen cells from mice immunized with sea nettle (Chrysaora quinquecirrha) venom exhibited a proliferative response after exposure to the homologous antigen or that of a related jellyfish, Physalia (Portuguese man-o'-war). Native venom was a more effective stimulant than heated, non-lethal venom. Ultraviolet light treatments administered to the skin either before or after venom sensitization suppressed the proliferative response of these internal immune cells.

Effects of verapamil and CSL antivenom on Chironex fleckeri (box-jellyfish) induced mortality.

Ovine antivenom prolonged survival in mice challenged with intravenous Chironex fleckeri venom over a limited dose range. Verapamil enhanced the beneficial effect of ovine antivenom. This data suggests that there may be an, as yet undefined, optimal antivenom dose for humans and that verapamil, used in combination with antivenom is an effective agent.