bluuebunny/crossref_metadata_2025 huggingface dataset:
Improved efficacy and tolerability of retinoic acid in acne vulgaris: a new topical formulation with cyclodextrin complex ψ
Authors: Anadolu RY , Sen T , Tarimci N , Birol A , Erdem C
Published: June 2004

Objectives  Retinoic acid (RA) has long been used, both topically and systemically, for disorders of keratinization, acne and related disorders. In the present study, the efficacy and tolerability of topical RA prepared as a cyclodextrin beta complex (β‐CD) is investigated in 66 acne vulgaris patients.

Methods  This randomized, double‐blind, placebo‐controlled study compares nightly topical application of RA/β‐CD complex hydrogel formulation (0.025%), RA/β‐CD complex in moisturizing base (0.025%), hydrogel base, moisturizer base or a commercial RA gel (0.05%) in acne vulgaris patients. Improvement of acne was assessed using a 5‐point improvement scale and by measuring sebum and moisture content of the skin using an SM 810 sebumeter/corneometer.

Results  After 3 months of treatment, mean scores of acne improvement on the 5‐point scale were 4 with the RA/β‐CD complex hydrogel formulation, 4.1 with the RA/β‐CD complex in moisturizing base, 1.2 with hydrogel placebo base, 1.1 with moisturizer placebo base and 3 with the commercial RA product. All patients treated with the commercial product experienced local side‐effects. One patient discontinued due to severe irritation. None of the patients treated with the RA/β‐CD complex in the moisturizing base and hydrogel formulation experienced significant local irritation, although the sebum content of the skin decreased after application of the RA/β‐CD preparations. This change was not significant compared to controls. The moisture content of the skin was better preserved in the group treated with the RA/β‐CD complex in the moisturizing base.

Conclusion  The topical RA/β‐CD complex, in hydrogel and moisturizing base, was more effective than the twice concentrated commercial RA product. There were few topical side‐effects with this new formulation, which increases patient compliance. Topical RA/β‐CD (0.025% RA) did not significantly reduce sebum secretion but may help to preserve optimum epidermal moisture content with the proper base formulation. This is the first study in the literature reporting efficacy and tolerability of the topical RA/β‐CD complex in acne vulgaris. We conclude that the topical RA/β‐CD complex displays an improved efficacy and tolerability profile and is an effective treatment alternative for acne vulgaris.
Treatment of papuloerythroderma of Ofuji with Re‐PUVA: a case report and review of the therapy§
Authors: Mutluer S , Yerebakan O , Alpsoy E , Ciftcioglu MA , Yilmaz E
Published: June 2004

Papuloerythroderma of Ofuji (PEO) is a disease of elderly men, characterized by intensely pruritic and widespread, red, flat‐topped papules with sparing of the body folds and creases (the so‐called ‘deck‐chair’ sign). The etiopathogenesis of the disease remains unknown. Psoralen plus UVA (PUVA), topical and systemic corticosteroids, etretinate, cyclosporin and interferon are the main approaches in the treatment of this rare disease. A case of PEO in a 60‐year‐old man who responded to retinoid plus PUVA (Re‐PUVA) treatment is reported here and a review of the therapy with other relevant cases is presented.
Evaluation of the safety of a non‐animal stabilized hyaluronic acid (NASHA – Q‐Medical, Sweden) in European countries: a retrospective study from 1997 to 2001
Authors: André P
Published: June 2004

Background  In Europe, several filler devices are currently on the market for use in aesthetic dermatology and some of them cause severe, permanent, adverse reactions. Since 1996 a non‐animal stabilized hyaluronic acid (NASHA) from Q‐Medical, Sweden, has been introduced and is becoming a leading product in aesthetic dermatology. Hyaluronic acid has no species specificity and skin testing is not recommended before treatment.

Objective  Our purpose was to evaluate the incidence of adverse reactions from 1997 to 2001 and the safety of NASHA after injections into the skin for aesthetic reasons.

Method  Surveys were sent to physicians in European countries that agreed to participate. This is a retrospective study. A total of 12 344 syringes were sold by the Q‐Medical to these physicians and we evaluated the total number of patients treated to 35% of this number (4320). We separated immediate hypersensitivity reactions from delayed reactions and analysed infectious and other types of reactions.

Results  From 1997 until 2001, 34 cases of hypersensitivity were reported: 16 cases of immediate hypersensitivity and 18 cases of delayed. The global risk of sensitivity is 0.8%. Since 2000, the amount of protein in the raw product has decreased and the incidence of hypersensitivity reactions is around 0.6%. As 50% of these reactions are immediate and resolved within less than 3 weeks, the risk of strong but transient, delayed reaction is around 0.3%. Four cases of abscess were reported. They were all sterile. No bacterial infection was found. Herpetic recurrence is possible after lip augmentation according to the technique of injection. No systemic reactions were reported.

Conclusion  NASHA is a very useful and safe filler product. Skin testing does not seem to be necessary.
Intramuscular bipenicillin vs. intravenous penicillin in the treatment of erysipelas in adults: randomized controlled study
Authors: Zeglaoui F , Dziri C , Mokhtar I , Ezzine N , Kharfi M , Zghal M , Fazaa B , Kamoun MR
Published: June 2004

The objective of the study was to evaluate the efficacy of intramuscular penicillin: mixture of benzyl penicillin and procain penicillin (2 MU × 2 times daily) and intravenous benzyl penicillin (4 MU × 6 times daily) in the treatment of hospitalized adult patients with erysipelas.

A prospective randomized unicentric trial was conducted. In total, 112 patients entered the study; 57 in the intramuscular group and 55 patients in the intravenous group completed the trial.

The failure rate was 14% for intramuscular group and 20% for the intravenous group (P = 0.40). Local complications such as of the leg abscesses were observed in the two groups (intravenous 9.1%, intramuscular 7%;P = 0477). Of the patients treated with intravenous benzyl penicillin, 25.5% presented complications related to the route (venitis). Intramuscular penicillin should be considered an effective and well‐tolerated treatment of erysipelas in adult patients.
Otosyphilis: report on six cases
Authors: Klemm E , Wollina U
Published: June 2004

Syphilis is a multisystemic disease that may also affect the inner ear. The objective of this study was to investigate the frequency, clinical presentation and outcome of otosyphilis. A retrospective analysis of inpatients seen by a team of ENT specialists and dermatologists at a teaching hospital in Dresden, Germany, during 1986–2000. Six cases of otosyphilis had been diagnosed. None was related to risk factors such as HIV infection or drug abuse. We identified four female and two male patients, age range 25–58 years. Four patients suffered from syphilis stage II, one from syphilis stage and another from congential syphilis. ENT complaints included loss or impairment of hearing, tinnitus and nystagmus. All were treated with penicillin G. Adjuvant therapy included systemic corticosteroids oral pentoxifillin in two patients each. Improvement of ENT symptoms was achieved in all but one patient, who suffered from congential syphilis. This study confirms that otosyphilis is still a complication of syphilis in developed countries. The first line therapy is systemic penicillin G. There is a lack of controlled trials for adjuvant treatments such as corticosteroids and pentoxifillin. Clinical cooperation between dermatologists/venereologists and ENT specialists supports the best outcome.
Elastophagocytosis: a feature of resolving Sweet's syndrome
Authors: Loo WJ , Rytina E , Banfield C
Published: June 2004

We report the case of a patient with atypical Sweet's syndrome characterized by an annular erythema that showed consumption of elastic fibres by giant cells and histiocytes. Although the lesions were found on sun‐exposed sites and the first biopsy demonstrated extensive elastophagocytosis, our patient did not have photodamaged skin clinically. A repeat biopsy 5 weeks later demonstrated an abundant collection of neutrophils supporting the diagnosis of Sweet's syndrome. To our knowledge, an elastolytic granulomatous reaction pattern has not been previously reported in Sweet's syndrome.
Graham Little–Piccardi–Lassueur syndrome associated with androgen insensitivity syndrome (testicular feminization)
Authors: Vega Gutiérrez J , Miranda‐Romero A , Pérez Milán F , Martínez García G
Published: June 2004

Graham Little–Piccardi–Lassueur syndrome is characterized by the presence of cicatricial alopecia on the scalp, keratosis pilaris in the skin of trunk and extremities, and non‐cicatricial hair loss in pubis and axillae. A frequent form of male pseudohermaphroditism is complete androgen insensitivity syndrome (CAIS), also known as testicular feminization syndrome. It refers to genetic males with XY karyotype who, owing to a lack of sensitivity in the peripheral androgenic receptors, develop a female phenotype. Axillary and pubic hair is typically scarce or absent. To our knowledge, this is the first case describing the association of the two processes. The presence of both processes in the same patient furthers our understanding of Graham Little–Piccardi–Lassueur syndrome as it rejects the influence of androgens in the alopecias accompanying this syndrome. The coincidence of non‐cicatricial alopecia in axillary and pubic hair in both processes is also remarkable.
Disease‐specific quality of life is associated with anxiety and depression in patients with acne
Authors: Yazici K , Baz K , Yazici AE , Köktürk A , Tot S , Demirseren D , Buturak V
Published: June 2004

Objective  We aimed to evaluate the relationships between acne severity, anxiety, depression and disease‐specific quality of life in patients with acne.

Method  A total of 61 patients with acne vulgaris and 38 healthy volunteers were included in the study. Acne severity was assessed by the Global Acne Grading System. All patients were asked to complete the Acne Quality of Life Scale (AQOL), Dermatology Life Quality Index (DLQI) and Hospital Anxiety and Depression Scale (HAD), and healthy controls to fill only the HAD.

Results  The mean HAD anxiety subscale (HAD‐A) and HAD depression subscale (HAD‐D) scores of the patients were significantly higher than those of the controls. The rates of subjects at risk for anxiety (26.2%) and for depression (29.5%) were significantly higher in the patient group than in the control group (0% and 7.9%, respectively). We found no correlations between acne severity and scores of AQOL, DLQI, HAD‐A and HAD‐D. AQOL and DLQI scores were positively correlated with HAD‐A and HAD‐D scores in the patient group. The patients at risk for anxiety had significantly higher scores on AQOL and DLQI compared to those who were not at risk. There were no statistically significant differences between the female and male patients with respect to AQOL, DLQI, HAD‐A and HAD‐D scores.

Conclusion  (1) Irrespective of the degree of severity, patients with acne are at increased risk for anxiety and depression compared to the normal population. (2) Acne negatively affects quality of life, and the greater the impairment of quality of life due to acne, the greater the level of anxiety and depression. (3) A greater impairment of dermatologic quality of life seems to put the patient at an increased risk for anxiety disorder.
Antitumour activity of paclitaxel and interferon‐alpha in a case of metastatic eccrine porocarcinoma
Authors: Gutermuth J , Audring H , Voit C , Trefzer U , Haas N
Published: June 2004

Background  Eccrine porocarcinoma (EP) is a rare malignant tumour arising in the acrosyringium, with about 50% of the cases developing local recurrence or metastatic disease. No standard therapy protocols for metastatic disease exist. In the past, only short remissions were achieved by applying combinations of cytotoxic agents, which were associated with severe side‐effects.

Aim of the study  In the case reported here, the aim was to find a protocol with fewer side‐effects for a patient who was not willing to undergo extensive polychemotherapy.

Subject  A 67‐year‐old male patient with local recurrence and regional lymph node metastases after resection of EP was treated with a combination of interferon‐alpha (IFN‐α) 9 million units s.c. three times per week and paclitaxel 100 mg/m2 weekly i.v., which shows a side‐effect profile similar to taxotere and is used in the treatment of a variety of neoplasms such as advanced squamous cell carcinoma.

Main outcome  This less aggressive treatment was tolerated well and the patient responded with minor remission and long‐term stable disease.
Pemphigus vulgaris induced by radiotherapy
Authors: Orion E , Matz H , Wolf R
Published: June 2004

Pemphigus can be triggered or induced by many insults, such as drugs, infections, viruses and neoplasms. X‐ray‐induced pemphigus has been recorded only rarely in the literature. We describe the case of a woman with pemphigus vulgaris in remission, who relapsed 1 week after completion of an irradiation course for breast cancer, in the exact location of the radiation. We review the previous 15 cases from the literature and outline the common features of those cases.
Immune response in demodicosis
Authors: Akilov OE , Mumcuoglu KY
Published: June 2004

Background  Demodex folliculorum and Demodex brevis are obligatory parasites in the hair follicles and in the pilosebaceous glands. Although most people are infested with these mites, only a small number develop the clinical symptoms of demodicosis. The objective of this study was to determine the distinguishing features of the immune response to the infestation of the skin by Demodex mites.

Methods  Twenty‐nine patients with human demodicosis and 13 age‐ and sex‐matched healthy subjects participated in the study. The presence of mites was determined by microscopic inspection of secretion from sebum glands. The immune response was evaluated in the peripheral blood by identifying membrane markers of different immune cells using monoclonal antibodies, while the concentration of immunoglobulin (Ig)A, IgM and IgG was calculated by simple radial immunodiffusion using anti‐IgA, anti‐IgM and anti‐IgG. The level of circulating immune complexes and total haemolytic complement, as well as the preparatory and digestive function of neutrophils, and the functional activity of leucocytes were also studied.

Results  The absolute number of CD95+ was higher in patients with demodicosis. The absolute number of CD3+, CD4+, CD8+ and CD16+ cells, the ratio CD3+/CD20+ and the functional activity of leucocytes were significantly lower in individuals infested with Demodex mites. No significant differences were found in the percentage and absolute number of CD20+ cells, the ratio of CD4+/CD8+ T‐cell subpopulations, circulating immune complexes, level of serum complement activity (CH50), activity and index of phagocytosis and the levels of IgA, IgM and IgG antibodies between individuals infested with Demodex mites and the control group.

Conclusion  The readiness of lymphocytes to undergo apoptosis increases in parallel to the increasing density of the mites. This could be the result of local immunosuppression caused by the mites, which allows them to survive in the host skin.
The sources of political orientations in post‐industrial society: social class and education revisited
Authors: Werfhorst Herman G. Van de , Graaf Nan Dirk de
Published: June 2004

This paper studies the impact of social class and education on political orientation. We distinguish the ‘old’ middle class from a new class of social/cultural specialists. However, the difference in their political orientation may especially be related to the level and field of education; the new middle class is more highly educated and often in fields of study that extensively address social competencies, characteristics independently affecting political outcomes. Analyses on Dutch data showed that education is more important in the prediction of ‘cultural’ liberal issues than social class. Economically‐oriented issues are more strongly affected by social class. This means that interests of the new middle class are served by liberal standpoints relating to a strong government and income redistribution policies, but not relating to cultural issues.
Actinic cheilitis: clinical and pathologic characteristics in 65 cases
Authors: Markopoulos A , Albanidou‐Farmaki E , Kayavis I
Published: June 2004

Objective:  The purpose of this study was to determine the clinical and histopathologic presentation of actinic cheilitis.

Study design:  A retrospective study on 65 patients attending an Oral Medicine clinic in Greece over a 10 year period. For each case the demographic, clinical and histopathologic information were evaluated.

Results:  The mean age at the time of diagnosis was 53.1 ± 11.4 years. Thirty‐nine patients (60%) used tobacco in any form. An outdoor occupation was indicated for 43 (66.2%) patients. The location of the lesions of actinic cheilitis was in all cases on the lower lip. Actinic cheilitis appeared in three forms; white non‐ulcerated lesions (29%), erosions or ulcers of the lip (48%), mixed white and erosive (23%). The histopathologic characteristics included increased thickness of keratin layer, alterations of the thickness of spinous cell layer, epithelial dysplasia, connective tissue changes, perivascular inflammation and basophilic changes of connective tissue. In 11 cases (16.9%) the presence of squamous cell carcinoma was observed.

Conclusions:  This case‐series highlights varied clinical presentation of actinic cheilitis among whom a high proportion developed squamous cell carcinoma.
Fungicidal effect of three new synthetic cationic peptides against Candida albicans
Authors: Nikawa H , Fukushima H , Makihira S , Hamada T , Samaranayake LP
Published: June 2004

Objective:  Peptide antibiotics are considered a new class of antifungal agents. Of these, an α‐helical, cationic peptide termed Dhvar 4, a relative of salivary histatin has been shown to be an antifungal of relatively high potency. Similarly, lactoferricin B (LFB) and a derivative thereof, LFB(17–30), disrupts the fungal cell membrane and acts against Candida albicans. As Dhvar 4 and LFB(17–30), exhibit almost identical amino acid sequences at their C‐terminal, we hypothesized that laboratory synthesis of peptides with an α‐helical structure and having similar amphipathic properties could lead to products with candidacidal activity. Hence, three such peptides – JH8194, JH8195 and JH 8944, were synthesized and their antifungal properties compared with recognized antifungals LFB, LFB(17–30), human lactoferricin (LFH), Histatin‐5 and Dhvar 4, against two isolates of C. albicans.

Materials and methods:  The antifungal agents were synthesized and their secondary structures evaluated according to a previously described protocol of Situ and Bobek (2000)Antimicrob Agents Chemother44: 1485–1493. The C. albicans strains were oral isolates from a human immunodeficiency virus‐infected (isolate A2) and a healthy (A6) individual. A standard concentration of yeasts was exposed to a range of dilutions of the agents for a specific duration and the cell death (viability) in terms of the resultant colony forming units  ml−1 was quantified.

Results:  Dhvar 4, showed the most α‐helical propensity, and was the least fungicidal while LFB and LFB(17–30) showed the highest antifungal potential, and demonstrated total kill of A6, and A2 at 5 and 10 μM concentrations, respectively whilst LFH killed both isolates at a l0 μM concentration. Of the three new synthetic peptides, JH 8194 was the most potent (total kill of A6/A2 strains at 1.25/2.5 μM), followed by JH 8195 (total kill of A6/A2 strains at 5/10 μM while JH 8944 was the least potent as a 25 μM concentration was required to kill either strain of Candida. On further analyses of the relationship between pI value of the peptides and their anticandicidal activity, a significant positive correlation was noted. In order to rule out a cytotoxic effect of the new synthetic peptides we compared the fungicidal and hemolytic activities under similar incubation conditions using freshly isolated erythrocytes and all three peptides exhibited no detectable hemolysis upto an concentration of 100 μM in contrast to the polyene antifungal amphotericin B that elicited significant initiation of hemolysis at a concentration of 5.0 μM.

Conclusion:  Our data suggest that laboratory synthesis of agents with an α‐helical structure and having amphipathic properties similar to known, natural antifungal agents may be a promising avenue to generate products with improved antifungal activity.
Effect of bone marrow transplantation on the immunolocalization of p53, hMSH2, and hMLH1 proteins on oral mucosa
Authors: Gomez RS , Pimenta FJGS , Guimarães ALS , Souza LN , Salomão UE , De Almeida HC , Vaz RR
Published: June 2004

Objective:  Considering that hMSH2, hMLH1 and p53 are important in maintaining genomic stability of the oral mucosa epithelium, the purpose of the present study was to investigate the immunolocalization of these proteins in the epithelium of the oral mucosa of patients submitted to bone marrow transplantation (BMT) compared with controls.

Materials and methods:  Twenty‐one samples of lip biopsies from BMT recipients were retrieved. Twenty samples of normal lower labial mucosa associated with mucocele in non‐transplanted patients were included as control group. The streptavidin–biotin complex stain was used to detect the human DNA mismatch repair proteins hMSH2, hMLH1 and p53 protein.

Results:  The main findings demonstrated that the mean number of suprabasal epithelial cells positive for MSH2 was statistically higher than the control group. The immunostaining of hMLH1 and p53 at the basal and suprabasal epithelial layers were statistically higher in the oral labial mucosa of the BMT patients compared with controls.

Conclusion:  The present study shows that oral epithelial cells of BMT patients show increased immunolocalization of the DNA repair related proteins.
Metal Binding by the D,DX
35
E Motif of Human Immunodeficiency Virus Type 1 Integrase: Selective Rescue of Cys Substitutions by Mn
2+
In Vitro
Authors: Gao Kui , Wong Steven , Bushman Frederic
Published: June 2004

The D,DX
35
E motif characteristic of retroviral integrase enzymes (INs) is expected to bind the required metal cofactors (Mg
2+
or Mn
2+
), but direct evidence for a catalytic role has been lacking. Here we used a metal rescue strategy to investigate metal binding. We established conditions for analysis of an activity of IN, disintegration, in both Mg
2+
and Mn
2+
, and tested IN mutants with cysteine substitutions in each acidic residue of the D,DX
35
E motif. Mn
2+
but not Mg
2+
can bind tightly to Cys, so if metal binding at the acidic residues is mechanistically important, it is expected that the Cys-substituted enzymes would be active in the presence of Mn
2+
only. Of the three acidic residues, a strong metal rescue effect was obtained for D116C, a weaker rescue was seen for D64C, and no rescue was seen with E152C. Modest rescue could also be detected for D116C in normal integration in vitro. Comparison to Ser and Ala substitutions at D116 established that the rescue was selective for Cys. Further studies of the response to pH suggest that the metal cofactor may stabilize the deprotonated nucleophile active in catalysis, and studies of the response to NaCl titrations disclose an additional role for the metal cofactor in stabilizing the IN-DNA complex.
Tax Relieves Transcriptional Repression by Promoting Histone Deacetylase 1 Release from the Human T-Cell Leukemia Virus Type 1 Long Terminal Repeat
Authors: Lu Hanxin , Pise-Masison Cynthia A. , Linton Rebecca , Park Hyeon Ung , Schiltz R. Louis , Sartorelli Vittorio , Brady John N.
Published: June 2004

Expression of human T-cell leukemia virus type 1 (HTLV-1) is regulated by the viral transcriptional activator Tax. Tax activates viral transcription through interaction with the cellular transcription factor CREB and the coactivators CBP/p300. In this study, we have analyzed the role of histone deacetylase 1 (HDAC1) on HTLV-1 gene expression from an integrated template. First we show that trichostatin A, an HDAC inhibitor, enhances Tax expression in HTLV-1-transformed cells. Second, using a cell line containing a single-copy HTLV-1 long terminal repeat, we demonstrate that overexpression of HDAC1 represses Tax transactivation. Furthermore, a chromatin immunoprecipitation assay allowed us to analyze the interaction of transcription factors, coactivators, and HDACs with the basal and activated HTLV-1 promoter. We demonstrate that HDAC1 is associated with the inactive, but not the Tax-transactivated, HTLV-1 promoter. In vitro and in vivo glutathione
S
-transferase-Tax pull-down and coimmunoprecipitation experiments demonstrated that there is a direct physical association between Tax and HDAC1. Importantly, biotinylated chromatin pull-down assays demonstrated that Tax inhibits and/or dissociates the binding of HDAC1 to the HTLV-1 promoter. Our results provide evidence that Tax interacts directly with HDAC1 and regulates binding of the repressor to the HTLV-1 promoter.
Geminate Structures of
African Cassava Mosaic Virus
Authors: Böttcher Bettina , Unseld Sigrid , Ceulemans Hugo , Russell Robert B. , Jeske Holger
Published: June 2004

Two types of geminate structures were purified from
African cassava mosaic geminivirus
(ACMV)-infected
Nicotiana benthamiana
plants and analyzed by electron cryomicroscopy and image reconstruction. After cesium sulfate density gradient centrifugation, they were separated into lighter top (T) and heavier bottom (B) components. T particles comigrated with host proteins, whereas B particles were concentrated in a cesium density typical for complete virions. Both particles were composed of two incomplete icosahedra of 11 capsomers each, but T particles were slightly larger (diameter, 22.5 nm) and less dense in the interior than B particles (diameter, 21.5 nm). T particles were frequently associated with small globules of ∼14 nm diameter of unknown origin. The overall structure of ACMV, a begomovirus transmitted by whiteflies, was similar to that of
Maize streak virus
(MSV), a mastrevirus transmitted by leafhoppers, although the vertices of the icosahedra were less pronounced. Models of ACMV coat proteins based on
Satellite tobacco necrosis virus
support the exposure of parts of the molecule essential for transmission specificity by whiteflies and provide possible structural explanations for the smaller protrusion of the ACMV capsid relative to MSV. The differences of ACMV and MSV virion shapes are discussed with reference to their different animal vectors.
P74 Mediates Specific Binding of
Autographa californica
M Nucleopolyhedrovirus Occlusion-Derived Virus to Primary Cellular Targets in the Midgut Epithelia of
Heliothis virescens
Larvae
Authors: Haas-Stapleton Eric J. , Washburn Jan O. , Volkman Loy E.
Published: June 2004

P74, an envelope protein of the occlusion-derived virus (ODV) of
Autographa californica
M nucleopolyhedrovirus (AcMNPV), is critical for oral infection of
Trichoplusia ni
larvae. The role of P74 during primary infection, however, is unknown. Here we provide evidence that P74 facilitates binding of AcMNPV ODV to a specific receptor within the larval midgut epithelia of another host species,
Heliothis virescens
. We adapted a fluorescence dequenching assay to compare binding, fusion, and competition of wild-type AcMNPV ODV in vivo with itself and with the ODV of a
p74
-deficient AcMNPV mutant. We found that relative to wild-type ODV, binding and fusion of ODV deficient in P74 were both qualitatively and quantitatively different. Unlike wild-type ODV, an excess of P74-deficient ODV failed to compete effectively with wild-type ODV binding, and the overall binding level of the mutant ODV was one-third that of the wild type. These results implicated P74 as an ODV attachment protein that binds to a specific receptor on primary target cells within the midgut.
Prion Infection of Skeletal Muscle Cells and Papillae in the Tongue
Authors: Mulcahy Ellyn R. , Bartz Jason C. , Kincaid Anthony E. , Bessen Richard A.
Published: June 2004

The presence of the prion agent in skeletal muscle is thought to be due to the infection of nerve fibers located within the muscle. We report here that the pathological isoform of the prion protein, PrP
Sc
, accumulates within skeletal muscle cells, in addition to axons, in the tongue of hamsters following intralingual and intracerebral inoculation of the HY strain of the transmissible mink encephalopathy agent. Localization of PrP
Sc
to the neuromuscular junction suggests that this synapse is a site for prion agent spread between motor axon terminals and muscle cells. Following intracerebral inoculation, the majority of PrP
Sc
in the tongue was found in the lamina propria, where it was associated with sensory nerve fibers in the core of the lingual papillae. PrP
Sc
staining was also identified in the stratified squamous epithelium of the lingual mucosa. These findings indicate that prion infection of skeletal muscle cells and the epithelial layer in the tongue can be established following the spread of the prion agent from nerve terminals and/or axons that innervate the tongue. Our data suggest that ingestion of meat products containing prion-infected tongue could result in human exposure to the prion agent, while sloughing of prion-infected epithelial cells at the mucosal surface of the tongue could be a mechanism for prion agent shedding and subsequent prion transmission in animals.
Human Immunodeficiency Virus Type 1 Tat Increases the Expression of Cleavage and Polyadenylation Specificity Factor 73-Kilodalton Subunit Modulating Cellular and Viral Expression
Authors: Calzado Marco A. , Sancho Rocío , Muñoz Eduardo
Published: June 2004

The human immunodeficiency virus type 1 (HIV-1) Tat protein, which is essential for HIV gene expression and viral replication, is known to mediate pleiotropic effects on various cell functions. For instance, Tat protein is able to regulate the rate of transcription of host cellular genes and to interact with the signaling machinery, leading to cellular dysfunction. To study the effect that HIV-1 Tat exerts on the host cell, we identified several genes that were up- or down-regulated intat-expressing cell lines by using the differential display method. HIV-1 Tat specifically increases the expression of the cleavage and polyadenylation specificity factor (CPSF) 73-kDa subunit (CPSF3) without affecting the expression of the 160- and 100-kDa subunits of the CPSF complex. This complex comprises four subunits and has a key function in the 3′-end processing of pre-mRNAs by a coordinated interaction with other factors. CPSF3 overexpression experiments and knockdown of the endogenous CPSF3 by mRNA interference have shown that this subunit of the complex is an important regulatory protein for both viral and cellular gene expression. In addition to the known CPSF3 function in RNA polyadenylation, we also present evidence that this protein exerts transcriptional activities by repressing themdm2gene promoter. Thus, HIV-1-Tat up-regulation of CPSF3 could represent a novel mechanism by which this virus increases mRNA processing, causing an increase in both cell and viral gene expression.
Vaccinia Virus G1 Protein, a Predicted Metalloprotease, Is Essential for Morphogenesis of Infectious Virions but Not for Cleavage of Major Core Proteins
Authors: Ansarah-Sobrinho Camilo , Moss Bernard
Published: June 2004

Genes encoding orthologs of the vaccinia virus G1 protein are present in all poxviruses for which sequence information is available, yet neither the role of the protein nor its requirement for virus replication is known. G1 was predicted to be involved in the cleavage of core proteins, based on a transfection study and the presence of an HXXEH motif found in a subset of metallopeptidases. In the present study, we engineered a recombinant vaccinia virus containing a single copy of the G1L gene with a C-terminal epitope tag that is stringently regulated by the
Escherichia coli lac
repressor. In the absence of inducer, expression of G1 was repressed and virus replication was inhibited. Rescue of infectious virus was achieved by expression of wild-type G1 in
trans
, but not when the putative protease active site residues histidine-41, glutamate-44, or histidine-45 were mutated. Nevertheless, the synthesis and proteolytic processing of major core and membrane proteins appeared unaffected under nonpermissive conditions, distinguishing the phenotype of the G1L mutant from one in which the gene encoding the I7 protease was repressed. Noninfectious virus particles, assembled in the absence of inducer, did not attain the oval shape or characteristic core structure of mature virions. The polypeptide composition of these particles, however, closely resembled that of wild-type virus. Full-length and shorter forms of the G1 protein were found in the core fraction of virus particles assembled in the presence of inducer, suggesting that G1 is processed by self-cleavage or by another protease.
Nef Proteins from Simian Immunodeficiency Virus-Infected Chimpanzees Interact with p21-Activated Kinase 2 and Modulate Cell Surface Expression of Various Human Receptors
Authors: Kirchhoff Frank , Schindler Michael , Bailer Nicola , Renkema G. Herma , Saksela Kalle , Knoop Volker , Müller-Trutwin Michaela C. , Santiago Mario L. , Bibollet-Ruche Frederic , Dittmar Matthias T. , Heeney Jonathan L. , Hahn Beatrice H. , Münch Jan
Published: June 2004

The accessory Nef protein allows human immunodeficiency virus type 1 (HIV-1) to persist at high levels and to cause AIDS in infected humans. The function of HIV-1 group M subtype Bnefalleles has been extensively studied, and a variety of in vitro activities believed to be important for viral pathogenesis have been established. However, the function ofnefalleles derived from naturally simian immunodeficiency virus (SIV)-infected chimpanzees, the original host of HIV-1, or from the HIV-1 N and O groups resulting from independent zoonotic transmissions remains to be investigated. In the present study we demonstrate that SIVcpz and HIV-1 group N or Onefalleles down-modulate CD4, CD28, and class I or II MHC molecules and up-regulate surface expression of the invariant chain (Ii) associated with immature major histocompatibility complex (MHC) class II. Furthermore, the ability of Nef to interact with the p21-activated kinase 2 was generally conserved. The functional activity of HIV-1 group N and Onefgenes did not differ significantly from group Mnefalleles. However, SIVcpznefgenes as a group showed a 1.8- and 2.0-fold-higher activity in modulating CD28 (P= 0.0002) and Ii (P= 0.016) surface expression, respectively, but were 1.7-fold less active in down-regulating MHC class II molecules (P= 0.006) compared to HIV-1 Mnefgenes. Our finding that primary SIVcpznefalleles derived from naturally infected chimpanzees modulate the surface expression of various human cellular receptors involved in T-cell activation and antigen presentation suggests that functionalnefgenes helped the chimpanzee virus to persist efficiently in infected humans immediately after zoonotic transmission.
RNA Interference Targeting VP1 Inhibits Foot-and-Mouth Disease Virus Replication in BHK-21 Cells and Suckling Mice
Authors: Chen Weizao , Yan Weiyao , Du Qingyun , Fei Liang , Liu Mingqiu , Ni Zheng , Sheng Zutian , Zheng Zhaoxin
Published: June 2004

RNA interference (RNAi) is a powerful tool to silence gene expression posttranscriptionally. In this study, we evaluated the antiviral potential of small interfering RNA (siRNA) targeting VP1 of foot-and-mouth disease virus (FMDV), which is essential during the life cycle of the virus and plays a key role in virus attachment to susceptible cells. We investigated in vivo the inhibitory effect of VP1-specific siRNAs on FMDV replication in BHK-21 cells and suckling mice, a commonly used small animal model. The results showed that transfection of siRNA-expressing plasmids gave an 80 to 90% reduction in the expression of FMDV VP1 in BHK-21 cells. Moreover, BHK-21 cells transiently transfected with siRNA-expressing plasmids were specifically resistant to FMDV infection when exposed to 100 50% tissue culture infective doses of virus, and the antiviral effects extended to almost 48 h postinfection. Furthermore, subcutaneous injection of siRNA-expressing plasmids in the neck made suckling mice significantly less susceptible to FMDV. In conclusion, our data suggests that RNAi may provide a viable therapeutic approach to treat FMDV infection.
The Two Major Human Metapneumovirus Genetic Lineages Are Highly Related Antigenically, and the Fusion (F) Protein Is a Major Contributor to This Antigenic Relatedness
Authors: Skiadopoulos Mario H. , Biacchesi Stéphane , Buchholz Ursula J. , Riggs Jeffrey M. , Surman Sonja R. , Amaro-Carambot Emerito , McAuliffe Josephine M. , Elkins William R. , St. Claire Marisa , Collins Peter L. , Murphy Brian R.
Published: June 2004

The growth properties and antigenic relatedness of the CAN98-75 (CAN75) and the CAN97-83 (CAN83) human metapneumovirus (HMPV) strains, which represent the two distinct HMPV genetic lineages and exhibit 5 and 63% amino acid divergence in the fusion (F) and attachment (G) proteins, respectively, were investigated in vitro and in rodents and nonhuman primates. Both strains replicated to high titers (≥6.0 log
10
) in the upper respiratory tract of hamsters and to moderate titers (≥3.6 log
10
) in the lower respiratory tract. The two lineages exhibited 48% antigenic relatedness based on reciprocal cross-neutralization assay with postinfection hamster sera, and infection with each strain provided a high level of resistance to reinfection with the homologous or heterologous strain. Hamsters immunized with a recombinant human parainfluenza virus type 1 expressing the fusion F protein of the CAN83 strain developed a serum antibody response that efficiently neutralized virus from both lineages and were protected from challenge with either HMPV strain. This result indicates that the HMPV F protein is a major antigenic determinant that mediates extensive cross-lineage neutralization and protection. Both HMPV strains replicated to low titers in the upper and lower respiratory tracts of rhesus macaques but induced high levels of HMPV-neutralizing antibodies in serum effective against both lineages. The level of HMPV replication in chimpanzees was moderately higher, and infected animals developed mild colds. HMPV replicated the most efficiently in the respiratory tracts of African green monkeys, and the infected animals developed a high level of HMPV serum-neutralizing antibodies (1:500 to 1:1,000) effective against both lineages. Reciprocal cross-neutralization assays in which postinfection sera from all three primate species were used indicated that CAN75 and CAN83 are 64 to 99% related antigenically. HMPV-infected chimpanzees and African green monkeys were highly protected from challenge with the heterologous HMPV strain. Taken together, the results from hamsters and nonhuman primates support the conclusion that the two HMPV genetic lineages are highly related antigenically and are not distinct antigenic subtypes or subgroups as defined by reciprocal cross-neutralization in vitro.
Functional Analysis of the Putative Fusion Domain of the Baculovirus Envelope Fusion Protein F
Authors: Westenberg Marcel , Veenman Frank , Roode Els C. , Goldbach Rob W. , Vlak Just M. , Zuidema Douwe
Published: June 2004

Group II nucleopolyhedroviruses (NPVs), e.g.,
Spodoptera exigua M
NPV, lack a GP64-like protein that is present in group I NPVs but have an unrelated envelope fusion protein named F. In contrast to GP64, the F protein has to be activated by a posttranslational cleavage mechanism to become fusogenic. In several vertebrate viral fusion proteins, the cleavage activation generates a new N terminus which forms the so-called fusion peptide. This fusion peptide inserts in the cellular membrane, thereby facilitating apposition of the viral and cellular membrane upon sequential conformational changes of the fusion protein. A similar peptide has been identified in NPV F proteins at the N terminus of the large membrane-anchored subunit F
1
. The role of individual amino acids in this putative fusion peptide on viral infectivity and propagation was studied by mutagenesis. Mutant F proteins with single amino acid changes as well as an F protein with a deleted putative fusion peptide were introduced in
gp64
-null
Autographa californica M
NPV budded viruses (BVs). None of the mutations analyzed had an major effect on the processing and incorporation of F proteins in the envelope of BVs. Only two mutants, one with a substitution for a hydrophobic residue (F152R) and one with a deleted putative fusion peptide, were completely unable to rescue the
gp64
-null mutant. Several nonconservative substitutions for other hydrophobic residues and the conserved lysine residue had only an effect on viral infectivity. In contrast to what was expected from vertebrate virus fusion peptides, alanine substitutions for glycines did not show any effect.
Sequence and Organization of the
Neodiprion lecontei
Nucleopolyhedrovirus Genome
Authors: Lauzon Hilary A. M. , Lucarotti Christopher J. , Krell Peter J. , Feng Qili , Retnakaran Arthur , Arif Basil M.
Published: June 2004

All fully sequenced baculovirus genomes, with the exception of the dipteran
Culex nigripalpus
nucleopolyhedrovirus (CuniNPV), have previously been from Lepidoptera. This study reports the sequencing and characterization of a hymenopteran baculovirus,
Neodiprion lecontei
nucleopolyhedrovirus (NeleNPV), from the redheaded pine sawfly. NeleNPV has the smallest genome so far published (81,755 bp) and has a GC content of only 33.3%. It contains 89 potential open reading frames, 43 with baculovirus homologues, 6 identified by conserved domains, and 1 with homology to a densovirus structural protein. Average amino acid identity of homologues ranged from 19.7% with CuniNPV to 24.9% with
Spodoptera exigua
nucleopolyhedrovirus. The conserved set of baculovirus genes has dropped to 29, since NeleNPV lacks an F protein homologue (ac23/ld130). NeleNPV contains 12 conserved lepidopteran baculovirus genes, including that for DNA binding protein, late expression factor 11 (
lef
-
11
),
polyhedrin
,
occlusion derived virus envelope protein
-
18
(
odv-e18
),
p40
, and
p45
, but lacks 21 others, including
lef
-
3
,
me53
,
immediate early gene
-
1
,
lef
-
6
,
pp31
,
odv-e66
,
few polyhedra 25k
,
odv-e25
,
protein kinase
-
1
,
fibroblast growth factor
, and
ubiquitin
. The lack of identified baculovirus homologues may be due to difficulties in identification, differences in host-virus interactions, or other genes performing similar functions. Gene parity plots showed limited colinearity of NeleNPV with other baculoviruses, and phylogenetic analysis indicates that NeleNPV may have existed before the lepidopteran nucleopolyhedrovirus and granulovirus divergence. The creation of two new
Baculoviridae
genera to fit hymenopteran and dipteran baculoviruses may be necessary.
Noninfectious X4 but Not R5 Human Immunodeficiency Virus Type 1 Virions Inhibit Humoral Immune Responses in Human Lymphoid Tissue Ex Vivo
Authors: Fitzgerald Wendy , Sylwester Andrew W. , Grivel Jean-Charles , Lifson Jeffrey D. , Margolis Leonid B.
Published: June 2004

Ex vivo human immunodeficiency virus type 1 (HIV-1) infection of human lymphoid tissue recapitulates some aspects of in vivo HIV-1 infection, including a severe depletion of CD4+T cells and suppression of humoral immune responses to recall antigens or to polyclonal stimuli. These effects are induced by infection with X4 HIV-1 variants, whereas infection with R5 variants results in only mild depletion of CD4+T cells and no suppression of immune responses. To study the mechanisms of suppression of immune responses in this ex vivo system, we used aldrithiol-2 (AT-2)-inactivated virions that have functional envelope glycoproteins but are not infectious and do not deplete CD4+T cells in human lymphoid tissues ex vivo. Nevertheless, AT-2-inactivated X4 (but not R5) HIV-1 virions, even with only a brief exposure, inhibit antibody responses in human lymphoid tissue ex vivo, similarly to infectious virus. This phenomenon is mediated by soluble immunosuppressive factor(s) secreted by tissue exposed to virus.
Myxoma Virus M11L Prevents Apoptosis through Constitutive Interaction with Bak
Authors: Wang Gen , Barrett John W. , Nazarian Steven H. , Everett Helen , Gao Xiujuan , Bleackley Chris , Colwill Karen , Moran Michael F. , McFadden Grant
Published: June 2004

M11L, a 166-amino-acid antiapoptotic protein of myxoma virus, was previously shown to bind to the peripheral benzodiazepine receptor by hydrophobic interactions at the outer mitochondrial membrane. Here we demonstrate that an additional property of M11L is the ability to constitutively form inhibitory complexes with the proapoptotic Bcl-2 family member Bak in human cells. This binding interaction was identified by both FLAG-tagged pull-down assays and tandem affinity purification from transfected and virus-infected human cells. M11L binds constitutively to human Bak and, under some inducible conditions, to human Bax as well, but not to the other Bcl-2 family members (Bad, Bid, Bcl-2). When stably expressed in human embryonic kidney (HEK293) cells, M11L effectively protects these cells from Fas ligand-induced apoptosis, thereby blocking release of cytochrome
c
, activation of caspase 9, and cleavage of poly(ADP-ribose) polymerase. We also demonstrate in coexpression studies that M11L can interact with Bak independently of any involvement with Bax. Furthermore, cells stably expressing M11L function to prevent apoptosis that is induced by overexpression of Bak. We conclude that M11L inhibits, in a species-independent fashion, apoptotic signals mediated by activation of Bak.
Mutations in the Human Cytomegalovirus UL27 Gene That Confer Resistance to Maribavir
Authors: Chou Sunwen , Marousek Gail I. , Senters Anne E. , Davis Michelle G. , Biron Karen K.
Published: June 2004

Previous drug selection experiments resulted in the isolation of a human cytomegalovirus (CMV) UL97 phosphotransferase mutant resistant to the benzimidazole compound maribavir (1263W94), reflecting the anti-UL97 effect of this drug. Three other CMV strains were plaque purified during these experiments. These strains showed lower-grade resistance to maribavir than the UL97 mutant. Extensive DNA sequence analyses showed no changes from the baseline strain AD169 in UL97, the genes involved in DNA replication, and most structural proteins. However, changes were identified in UL27 where each strain contained a different mutation (R233S, W362R, or a combination of A406V and a stop at codon 415). The mutation at codon 415 is predicted to truncate the expressed UL27 protein by 193 codons (32% of UL27) with a loss of nuclear localization. The expression of full-length UL27 as a green fluorescent fusion protein in uninfected fibroblasts resulted in nuclear and nucleolar fluorescence, whereas cytoplasmic localization was observed when codons 1 to 415 were similarly expressed. Viable UL27 deletion mutants were created by recombination and showed slight growth attenuation and maribavir resistance in cell culture. Marker transfer experiments confirmed that UL27 mutations conferred maribavir resistance. The UL27 sequence was well conserved in a sample of 16 diverse clinical isolates. Mutation in UL27, a betaherpesvirus-specific early gene of unknown biological function, may adapt the virus for growth in the absence of UL97 activity.