First Report of Botrytis hyacinthi on Pineapple Lily in California.

Pineapple lily (Eucomis vandermerwei) is grown for its exotic-looking flower spike that looks like a small pineapple. It is used as an accent plant in the garden or for its long-lasting cut flowers. In late November 2007, plant foliage was submitted to the California State Diagnostic Laboratory by a commercial grower for phytosanitary inspection before shipment of bulbs to Europe. Leaves had wet, brown, elliptical spots of 1 to 2 cm long, some with yellow halos. The interior of the older spots was pale brown and papery. Conidiophores typical of Botrytis spp. were observed on the abaxial side of the leaf in the interior of the older spots. Isolations on half-strength acidified potato dextrose agar (APDA) yielded a grayish mycelium with conidiophores developing on the plant tissue only. The agar was grayish yellow, especially when viewed from the underside. Conidiophores measured 275 to 650 µm × 15 to 20 µm (411.2 × 15.5 µm average). Conidia were light brown, subglobose to broadly ellipsoidal, and measuring 10 to 15 × 13 to 20 µm (11.8 × 16.2 µm average). Scattered, black sclerotia measuring 335 to 1,007 × 518 to 1,079 µm (668.9 × 743.9 µm average) formed on the medium after approximately 7 days. Pathogenicity was confirmed by inoculating four Eucomis spp. plants with one inoculation per leaf, four leaves per plant. Each leaf was wounded with a sterile pushpin and three agar plugs from 4-day-old cultures were placed in a plastic screw-cap lid filled with sterile water and clipped onto each wound. Plants were misted with water, covered with a plastic bag, and placed in a growth chamber at 16°C (12-h photoperiod) for 48 h after which the agar plugs and caps were removed. An equal number of plants were wounded and mock inoculated with APDA. Pathogenicity experiments were repeated. After 14 days, all inoculated leaves had lesions and the fungus was reisolated. No Botrytis spp. were isolated from the mock-inoculated plants. Our sequence of the intergenic spacer regions of one isolate, GenBank FJ10809, matched Botrytis hyacinthi sequence AJ716297 with 99.8% identity. Using molecular (3) and morphological characters (1,4), the pathogen was identified as B. hyacinthi, the cause of fire disease in hyacinths. This pathogen was described previously on hyacinths in Washington State (2), the Netherlands (4), and the United Kingdom (2). The importance and economic impact of this disease appears to be limited because it has only been observed on mature or senescing foliage and not bulbs. To our knowledge, this is the first report of B. hyacinthi causing a leaf spot on pineapple lily. References: (1) C. J. Gould et al. Plant Dis. Rep. 42:534, 1958. (2) W. C. Moore. Page 14 in: Diseases of Bulbs. Bull. No. 117. Minist. Agric. Fish. London, 1939. (3) M. Staats et al. Mol. Biol. Evol. 22:333, 2005. (4) J. Westerdijk and F. H. Van Beyma Thoe Kingma. Meded. Phytopathol. Lab. "Willie Commelin Scholten" 12:15, 1928.

Genetic determinants of Venezuelan equine encephalitis emergence.

Following a period of inactivity from 1973-1991, Venezuelan equine encephalitis (VEE) reemerged during the past decade in South America and Mexico. Experimental studies of VEE virus (VEEV) infection of horses with virus strains isolated during these outbreaks have revealed considerable variation in the ability of equine-virulent, epizootic strains to exploit horses as efficient amplification hosts. Subtype IC strains from recent outbreaks in Venezuela and Colombia amplify efficiently in equines, with a correlation between maximum viremia titers and the extent of the outbreak from which the virus strain was isolated. Studies of enzootic VEEV strains that are believed to represent progenitors of the epizootic subtypes support the hypothesis that adaptation to efficient replication in equines is a major determinant of emergence and the ability of VEEV to spread geographically. Correlations between the ability of enzootic and epizootic VEEV strains to infect abundant, equiphilic mosquitoes, and the location and extent of these outbreaks, also suggest that specific adaptation to Ochlerotatus taeniorhynchus mosquitoes is a determinant of some but not all emergence events. Genetic studies imply that mutations in the E2 envelope glycoprotein gene are major determinants of adaptation to both equines and mosquito vectors.

Recombinant sindbis/Venezuelan equine encephalitis virus is highly attenuated and immunogenic

Venezuelan equine encephalitis virus (VEEV) is an important, naturally emerging zoonotic virus. VEEV was a significant human and equine pathogen for much of the past century, and recent outbreaks in Venezuela and Colombia (1995), with about 100,000 human cases, indicate that this virus still poses a serious public health threat. The live attenuated TC-83 vaccine strain of VEEV was developed in the 1960s using a traditional approach of serial passaging in tissue culture of the virulent Trinidad donkey (TrD) strain. This vaccine presents several problems, including adverse, sometimes severe reactions in many human vaccinees. The TC-83 strain also retains residual murine virulence and is lethal for suckling mice after intracerebral (i.c.) or subcutaneous (s.c.) inoculation. To overcome these negative effects, we developed a recombinant, chimeric Sindbis/VEE virus (SIN-83) that is more highly attenuated. The genome of this virus encoded the replicative enzymes and the cis-acting RNA elements derived from Sindbis virus (SINV), one of the least human-pathogenic alphaviruses. The structural proteins were derived from VEEV TC-83. The SIN-83 virus, which contained an additional adaptive mutation in the nsP2 gene, replicated efficiently in common cell lines and did not cause detectable disease in adult or suckling mice after either i.c. or s.c. inoculation. However, SIN-83-vaccinated mice were efficiently protected against challenge with pathogenic strains of VEEV. Our findings suggest that the use of the SINV genome as a vector for expression of structural proteins derived from more pathogenic, encephalitic alphaviruses is a promising strategy for alphavirus vaccine development.

Effect of reserpine pretreatment on avian erythrocyte carbonic anhydrase activation by isoproterenol.

We studied the action of beta-adrenergic agonists on Japanese quail erythrocyte carbonic anhydrase (CA) in vitro. Earlier we had reported that epinephrine increased CA activity by 14%; the present study focused on an attempt to increase the size of this response. Washed erythrocytes from reserpine-treated (1 mg/kg daily i.m. for 3 days) and control birds were incubated for 40 min in the presence of isoproterenol 10(-6) mol/l. The activity of CA expressed as Wilbur-Anderson units/mg hemoglobin was increased by as much as 42% in reserpine-treated birds over the control depending on the conditions. Addition of 10(-5) mol/l of the beta-adrenergic antagonist propranolol inhibited the isoproterenol-induced effect in nonreserpinized birds. We conclude that pretreatment with reserpine, which was accompanied by a fall in plasma catecholamine levels, particularly epinephrine levels, enhanced the activation of CA by isoproterenol.

Efficacy of ceftibuten for acute otitis media caused by Hemophilus influenzae: an animal study.

Ceftibuten is a new oral cephalosporin with an unusual stability to beta-lactamases that can hydrolyze other extended-spectrum cephalosporins. Using the chinchilla animal model, we compared the efficacy of ceftibuten (n = 33) with that of saline (n = 34), ampicillin (n = 32), and cefixime (n = 31) for the treatment of acute otitis media caused by beta-lactamase-producing nontypeable Hemophilus influenzae. Ceftibuten was superior to ampicillin regarding the time necessary to sterilize the middle ear (p < .001) and eliminate effusion (p < .001). The mean days of therapy required for bacteriologic cure were 2.57 for ceftibuten, 2.95 for cefixime, 7.95 for ampicillin, and 8.16 for saline. At the conclusion of therapy, chinchillas treated with ceftibuten had a significantly lower prevalence of positive cultures and middle ear effusion than did animals treated with ampicillin. No significant differences were observed between ceftibuten and cefixime. The results of this randomized, investigator-blinded experiment warrant further consideration of ceftibuten as a second-line agent for acute otitis media caused by ampicillin-resistant H influenzae.

Determination of acetazolamide in human serum by enzymatic assay.

Carbonic anhydrase (CA) inhibitors, such as acetazolamide (AZ), formerly used as diuretics, still play a role in the treatment of glaucoma, epilepsy, and altitude sickness. There is now hard evidence from both in vitro and in vivo studies in animals that carbonic anhydrase plays a vital function in bone loss. Acetazolamide blocks bone resorption in these experimental models. We have postulated that acetazolamide has potential for the treatment of human conditions associated with bone loss. In preparation for a clinical trial of acetazolamide's effectiveness in this regard, we developed an enzymatic method for determining the total concentration of acetazolamide in human serum. Acetazolamide is stripped from binding to serum proteins by adding 10(-6) M salicylic acid and adjusting the pH to 2.5, followed by ultrafiltration through a membrane (10 kD cutoff). The latter permits the free acetazolamide to enter the filtrate but retains any carbonic anhydrase (31 kD) which may contaminate the serum from hemolysis. The carbonic anhydrase inhibitory activity in the filtrate, representing the acetazolamide, is determined in a carbonic anhydrase assay using acetazolamide as the standard. Recoveries of acetazolamide added to human serum ranged from 83% to 94% depending on the concentration. Precision, as judged by the coefficient of variation, was 10.5%.

Carbonic anhydrase inhibition by flurbiprofen and related agents.

Flurbiprofen, a nonsteroidal anti-inflammatory drug (NSAID), reduces bone resorption in periodontal disease. This therapeutic effect has been attributed to inhibition of prostaglandin synthesis. In view of the importance of carbonic anhydrase (CA) in bone resorption, we examined the CA-inhibitory properties of flurbiprofen using bovine and human CA II and compared them with those of acetazolamide and two other NSAIDs, ibuprofen and indomethacin. Flurbiprofen inhibited both human and bovine erythrocyte CA II but to a much lesser degree than acetazolamide. Ibuprofen and indomethacin were much less active in inhibiting CA II than flurbiprofen.

Therapy for genital warts.

This article presents the spectrum of genital wart infection, including possible subclinical infection and reinfection from partners. Present therapy is effective in warts of recent onset but usually requires multiple treatments and is painful, and warts are frequently recurrent. Trials of combination therapy appear to have greater efficacy than does single-agent treatment. Problems for the future include the best management of warts in the sexually abused child, the immunocompromised host, and the pregnant patient. Genital warts are increasingly linked with the potential development of carcinoma, mandating more effective treatment regimens in the future.

Bacterial, viral, fungal, and parasitic infections in HIV disease and AIDS.

Infectious complications are the most common cutaneous manifestations of HIV infection. In this article, the bacterial, viral, fungal, and parasitic infections seen in HIV-infected patients are described and illustrated. The relation of these infections to the overall health of the patient and current therapies that have been found beneficial are outlined.

Merkel cell tumor of the thigh.

This case of a Merkel cell carcinoma is unusual due to the occurrence of the tumor on the thigh; most Merkel cell tumors have been found on the sun-exposed region of the head and neck. Histologically, the nodule was composed of sheets of uniform, poorly differentiated cells with a high nuclear to cytoplasmic ratio. Electron microscopy revealed perinuclear filaments, scattered dense core granules, and complex, interdigitating processes within cytoplasmic membranes. Treatment consisted of surgical excision of the tumor with a wide margin.

Cutaneous pneumocystosis.

A 42-year-old man with the acquired immunodeficiency syndrome (AIDS) presented with a 9-month history of progressive hearing loss and bilateral external auditory canal masses. Biopsy of the right ear mass detected Pneumocystis carinii. The patient was treated with a 7-day course of intravenous trimethoprim, followed by a 3-week course of oral trimethoprim-sulfamethoxazole, with a marked reduction in the size of his ear masses. This is the first reported case of skin involvement by P. carinii in a patient with AIDS. Because of our patient's documented response to treatment, we stress the importance of biopsy of cutaneous lesions in this group of patients, who may have a potentially treatable condition.

Primary cutaneous amyloidosis: identically distributed lesions in identical twins.

Male HLA-identical twins with numerous congenital abnormalities were discovered to have lichen amyloidosus in similar distribution on their chests and abdomens. Pruritus was absent in both. The finding of identical distribution of nonpruritic lesions was previously reported in familial lichen amyloidosus. This suggests that a subset of these patients have a nevoid condition in which a circumscribed patch of keratinocytes undergoes filamentous degeneration and may appear in identical distribution in family members in the absence of pruritus.

Amyloid deposition after psoriasis therapy with psoralen and long-wave ultraviolet light.

Histologic examination of skin from 52 patients under continued treatment with methoxsalen and long-wave ultraviolet irradiation (PUVA) disclosed small superficial dermal deposits in six subjects who had completed one year of therapy and in six subjects who had completed two years of therapy. These deposits, by light and electron microscopic criteria, are similar to those of lichen amyloidosus. Amyloid deposition should be regarded as a possible complication of PUVA therapy.