bluuebunny/crossref_metadata_2025 huggingface dataset:
No transmission of porcine endogenous retrovirus after transplantation of adult porcine islets into diabetic nude mice and immunosuppressed rats
Authors: Goto Masafumi , Maeda Akira , Elfman Lotta , Suling Kristen M. , Wood James C. , Patience Clive , Groth Carl G. , Wennberg Lars
Published: June 2004

Background:  The aim of this study was to investigate whether transmission of porcine endogenous retrovirus (PERV) occurs in a model of diabetes reversal by the xenotransplantation of adult porcine islets (APIs) into immunoincompetent diabetic rodents.

Methods:  Black‐6 nu/nu mice and Lewis rats were immunosuppressed with cyclosporin A (CsA) and FTY 720, and rendered diabetic with streptozotocin. Purified APIs were transplanted into the renal subcapsular space; 5000 islet equivalents (IEQs) were used in the nude mice (n = 4) and 40 000 IEQs in the rats (n = 4). The nude mice were sacrificed at 75 days after transplantation. In order to confirm chronic xenograft function, the graft‐bearing kidney was removed prior to sacrifice. The rats were followed until xenograft rejection, at which time they were sacrificed. Immediately after sacrifice, tissue samples (liver, spleen, and small intestine) were taken for analysis. Quantitative polymerase chain reaction (PCR) was used to assess evidence of PERV transmission, and porcine cell chimerism.

Results:  All animals became normoglycemic within 48 h of transplantation. The nude mice remained normoglycemic during the 75‐day study period, with removal of the graft‐bearing kidney resulting in prompt hyperglycemia. The rats remained normoglycemic until xenograft rejection, which occurred at 66 ± 28 days. Despite the evidence of porcine cell microchimerism in recipients, real‐time PCR detected no evidence of PERV transmission in any of the tissue specimens tested.

Conclusions:  There was no evidence of PERV transmission following transplantation of pig islets into diabetic nude mice and immunosuppressed rats.
MyD88‐dependent toll‐like receptor signalling is not a requirement for fetal islet xenograft rejection in mice
Authors: Schmidt Peter , Krook Henrik , Goto Masafumi , Korsgren Olle
Published: June 2004

Abstract:  Background:  Rejection of pancreatic islet xenografts in mice shares immunopathological features with a Th1‐associated delayed‐type hypersensitivity (DTH) reaction. The aim of the present study was to investigate the mechanism of acute cellular xenograft rejection in a strain of mice with a targeted gene disruption of the toll‐like receptor (TLR) signal adaptor protein MyD88. These mice have been shown to have markedly impaired Th1 immunity.

Methods:  The MyD88‐/‐ and normal mice were transplanted with 2 μl of fetal porcine islet‐like cell clusters (ICC) under the left kidney capsule. On days 3, 6 or 12 after transplantation the mice were killed and the grafts either prepared for immunohistochemistry or real‐time quantitative reverse transcriptase polymerase chain reaction (RT‐PCR). The number of remaining ICC and infiltrating cells with different phenotypic characteristics was assessed semi‐quantitatively. Grafts used for quantitative RT‐PCR were analysed for content of murine mRNA of interferon (IFN)‐γ, interleukin (IL)‐12p40, IL‐4 and IL‐10.

Results:  On day 3, the rejection process was initiated in both MyD88‐/‐ and normal mice as characterized by a moderate infiltration of F4/80+ and MAC‐1+ macrophages and occasional CD3+ and CD4+ cells. Expression of IFN‐γ and IL‐12p40 was lower but still detectable in the MyD88‐/‐ mice, when compared with control animals. By day 6, rejection was almost completed in all animals with only few ICC remaining. 12 days after transplantation all grafts were completely destroyed and heavily infiltrated by macrophages. Moderate numbers of CD3+ and CD4+ and occasional CD8+ cells were also present.

Conclusions:  Islet xenograft rejection was found to persist in MyD88‐/‐ mice. Despite a relatively lower expression of the Th1‐associated cytokines IFN‐γ and IL12‐p40 within the xenograft area, both the time course and morphological pattern of the rejection were essentially similar to that found in normal animals. Hence, MyD88‐dependent TLR signalling does not appear to be a crucial component of acute cellular xenograft rejection.
Bone marrow transplantation from α1,3‐galactosyltransferase gene‐knockout pigs in baboons
Authors: Tseng Y.‐L. , Dor F.J.M.F , Kuwaki K. , Ryan D. , Wood J. , Denaro M. , Giovino M. , Yamada K. , Hawley R. , Patience C. , Schuurman H.‐J. , Awwad M. , Sachs D.H. , Cooper D.K.C.
Published: June 2004

Abstract:  Background:  Successful hematopoietic cell allotransplantation results in donor‐specific tolerance, but this approach has been unsuccessful in the wild‐type pig‐to‐baboon xenotransplantation model, as pig cells were lost from the circulation within 5 days. However, after cessation of immunosuppressive therapy on day 28, all baboons demonstrated non‐specific unresponsiveness on mixed leukocyte reaction (MLR) for at least 30 days. We have now investigated the transplantation of bone marrow (BM) cells from miniature swine homozygous for α1,3‐galactosyltransferase gene‐knockout (GalT‐KO).

Methods:  Baboons (n = 3) were pre‐treated with whole body and thymic irradiation, anti‐thymocyte globulin, and splenectomy, and received immunosuppressive and supportive therapy for 28 days. BM was harvested from GalT‐KO swine (n = 3). The baboons were monitored for the presence of pig cells by flow cytometry and colony‐forming units (CFUs), and for cellular reactivity by MLR.

Results:  A mean of 11 × 108 BM cells/kg was infused into each baboon. The mean absolute numbers and percentages of pig cells detected in the blood at 2 h and on days 1, 2 and 4, respectively, were 641/μl (9.5%), 132/μl (3.4%), 242/μl (3.9%), and 156/μl (2.9%). One baboon died (from accidental hemorrhage) on day 6, at which time chimerism was present in the blood (2.0%) and BM (6.4%); pig cell engraftment in the BM was confirmed by polymerase chain reaction (PCR) of CFUs. In the two other baboons, blood chimerism was lost after day 5 but returned at low levels (<1%) between days 9 to 16 and 7 to 17, respectively, indicating transient BM engraftment. Both surviving baboons showed non‐specific unresponsiveness on MLR until they were euthanized on days 85 and 110, respectively.

Conclusions:  By using BM cells from GalT‐KO pigs, chimerism was detected at levels comparable with previous studies when 30‐fold more growth factor‐mobilized peripheral blood progenitor cells had been transplanted. In addition, cellular hyporesponsiveness was prolonged. However, long‐term engraftment and chimerism were not achieved.
Immune effect of hypertonic saline: fact or fiction?
Authors: Kølsen‐Petersen J. A.
Published: June 2004

Hypertonicity affects many parts of the immune system. Animal studies and experiments in isolated cell cultures show that hypertonicity reversibly suppresses several neutrophil functions and at the same time up‐regulates T‐lymphocyte function. Infusion of hypertonic saline with or without colloids may thus, besides providing efficient plasma volume expansion, ameliorate the detrimental consequences on the immune function of trauma, shock, reperfusion, and major surgery. However, the few clinical studies conducted to date, specifically addressing the immune effect of hypertonic saline infusion, have shown little, if any, effect on markers of immune function, and larger clinical trials have not demonstrated benefit in terms of morbidity or mortality. Thus, as opposed to animal and cell‐culture studies, the immune‐modulating properties of hypertonic saline infusion would appear to be of limited value in clinical practice.

This review presents in vitro studies, animal experiments, and clinical trials which investigated the consequences of hypertonic saline on markers of immune function.
Ketamine does not decrease postoperative pain after remifentanil‐based anaesthesia for tonsillectomy in adults
Authors: Van Elstraete A. C. , Lebrun T. , Sandefo I. , Polin B.
Published: June 2004

Background:  There are conflicting results concerning the pre‐emptive effect of ketamine on central sensitization following surgery. The aim of this prospective, randomized, double‐blind, placebo‐controlled study was to assess the effect of the N‐methyl‐D‐aspartate receptor antagonist ketamine on postoperative morphine consumption and pain score after remifentanil‐based anaesthesia in adult patients scheduled for tonsillectomy.

Methods:  We studied 40 adult patients undergoing elective tonsillectomy. Total intravenous anaesthesia was induced and maintained with remifentanil (0.125–1.0 µg kg−1 min−1) and propofol target‐controlled infusion. Patients in group K received a bolus dose of ketamine 0.5 mg kg−1 immediately after anaesthetic induction, followed by a continuous infusion of 2 µg kg−1 min−1. Saline was administered in the same sequence in group S. Propofol, remifentanil, and the study drug infusions were discontinued at the end of surgery.

Results:  Intraoperative remifentanil consumption (0.57 ± 0.18 in group K vs. 0.55 ± 0.14 µg kg−1 min−1 in group S), morphine requirement in the PACU (11 ± 3 in group K vs. 9 ± 4 mg in group S) and in the ward (22 ± 11 in group K vs. 25 ± 14 mg in group S), median time to first analgesia in the ward (338 ± 126 in group K vs. 328 ± 144 min in group S), and VAS pain scores were comparable in both groups.

Conclusion:  Small‐dose of ketamine does not seem to be a useful adjunct to remifentanil‐based anaesthesia during short, painful surgical procedures.
The effects of pre‐emptive epidural sufentanil on human immune function
Authors: Akural E. I. , Salomäki T. E. , Bloigu A. H. , Ryhänen P. , Tekay A. H. , Alahuhta S. M. , Surcel H.‐M.
Published: June 2004

Background:  Surgical stress and general anaesthesia suppress immune functions, including natural killer cell (NK) activity. This suppression could be attributed, at least in part, to the effect of opiates.

Methods:  Twenty patients undergoing abdominal hysterectomy received epidural sufentanil (50 μg) either before (pre‐emptive) or at the end (control group) of surgery. Post‐operative pain relief was provided using sufentanil from a patient‐controlled epidural analgesia (PCEA) system. Systemic immunity was assessed by determining leucocyte counts, NK cell counts and activity, lymphocyte response to mitogen stimulation, and secretion of pro‐inflammatory cytokines.

Results:  In the pre‐emptive group there was a significant decrease in NK activity on the first and third post‐operative day (P < 0.05) compared with baseline values and on the third postoperative day (P < 0.05) compared with the control group. The number of total leucocytes and neutrophiles increased in both groups post‐operatively, but no differences were found in the levels of mononuclear lymphocyte populations or in their mitogen responses. Interleukin‐6 (IL‐6) concentration increased in both groups after the operation. In addition, at the end of the surgery the IL‐6 level was greater in the control group than in the pre‐emptive group. Interleukin‐1 (IL‐1) levels had decreased significantly at the end of surgery and 4 h later compared with baseline levels in the pre‐emptive, but not in the control group.

Conclusions:  Pre‐emptive epidural sufentanil during combined propofol and isoflurane anaesthesia had minor effects on the immune response after hysterectomy. The lower production of pro‐inflammatory cytokines (IL‐1, IL‐6) in the pre‐emptive group compared with the control group is beneficial, but its clinical importance remains to be determined.
Predictive value of preoperative transthoracic echocardiography in patients undergoing adrenalectomy for pheochromocytoma
Authors: Devaux B. , Lentschener C. , Jude N. , Valensi L. , Pili‐Floury S. , Dousset B. , Ozier Y.
Published: June 2004

Background:  Adrenalectomy for pheochromocytoma is a life‐threatening procedure. Few echocardiographic assessments have been reported in patients undergoing adrenalectomy for pheochromocytoma.

Methods:  Sixty‐three consecutive patients undergoing adrenalectomy for pheochromocytoma underwent routine preoperative M‐mode and two‐dimensional echocardiography, and Doppler examination. Abnormal echocardiographic findings were defined as left ventricular dilatation or dysfunction (left ventricular percentage fractional shortening <30%), and/or left ventricular wall motion abnormalities, and/or left ventricular hypertrophy (left ventricular mass index >110 g m−2 in women and >134 g m−2 in men) and/or valvular abnormalities. Physical characteristics, daily urinary metanephrine and normetanephrine excretions, preoperative functional limitation, pre‐existing congestive heart failure, type and duration of surgery, and haemodynamic instability in the intra and postoperative periods were compared in patients with normal and abnormal echocardiographic findings.

Results:  Twenty‐four out of 63 patients were found to have abnormal preoperative echocardiography. There was no difference between patients with normal and abnormal preoperative echocardiography as regards to the investigated criteria, except for pre‐existing self‐reported functional limitation and chest pain suggesting coronary artery disease.

Conclusions:  The relevance of routine preoperative echocardiographic examination in patients scheduled for adrenalectomy for pheochromocytoma, who have no cardiac symptoms or clinical evidence of cardiac involvement, is questionable.
An Equal Distance? Individualisation, Gender and Intimacy in Distance Relationships
Authors: Holmes Mary , Why? Mary
Published: June 2004

Intimacy is usually thought to require physical proximity as a prerequisite, so how are intimate relationships maintained when partners do not live their daily lives in the same place? Couples who are frequently apart are not an entirely new phenomenon, but the dual-career, dual-household couples examined in this pilot study of contemporary distance relationships illustrate new ways of relating. This paper will explore these new ways of relating in terms of debates about the impact of individualisation processes on intimacy. It is argued that the extension of such processes to women has been limited. Ways of being intimate at a distance may offer alternative formulations of gender and power, but the rationalised timetabling necessary to maintain such relationships imposes new gendered constraints with bodily and emotional consequences.
Continuity and Change in Contemporary Ulster Protestantism*
Authors: Brewer John D.
Published: June 2004

This paper explores current trends in religious practice, observance and belief in Ulster Protestantism for elements of continuity and change. Using historical and survey data it is clear that there are strong elements of both. However, Protestant religiosity is not changing to the point that it constitutes secularisation, as sociologists of religion understand it. Nor are new trends in religiosity weakening ethno-national identities in Northern Ireland. This is because political identities are socially reproduced in ways that are independent of their religious roots and are thus unaffected by patterns of religiosity. Likely changes in Protestant religiosity in the future therefore offer no immediate panacea for altering the dynamics of Northern Irish politics.
Bayesian Analysis of Dynamic Magnetic Resonance Breast Images
Authors: de Pasquale Francesco , Barone Piero , Sebastiani Giovanni , Stander Julian
Published: June 2004

We describe an integrated methodology for analysing dynamic magnetic resonance images of the breast. The problems that motivate this methodology arise from a collaborative study with a tumour institute. The methods are developed within the Bayesian framework and comprise image restoration and classification steps. Two different approaches are proposed for the restoration. Bayesian inference is performed by means of Markov chain Monte Carlo algorithms. We make use of a Metropolis algorithm with a specially chosen proposal distribution that performs better than more commonly used proposals. The classification step is based on a few attribute images yielded by the restoration step that describe the essential features of the contrast agent variation over time. Procedures for hyperparameter estimation are provided, so making our method automatic. The results show the potential of the methodology to extract useful information from acquired dynamic magnetic resonance imaging data about tumour morphology and internal pathophysiological features.
Surrogate nits impregnated with white piedra – a case report
Authors: Ghorpade A
Published: June 2004

White piedral spores packed inside empty pedicular nits were accidentally found on microscopic examination in a 42‐year‐old Indian woman who presented with hair loss. The diagnosis of piedra was confirmed on culture. She responded to topical 2% miconazole nitrate solution and manual removal of the nits. This is the first case report of pedicular nits found to be impregnated with spores of white piedra.
HIV seronegative eosinophilic pustular folliculitis successfully treated with doxicycline
Authors: Brazzelli V , Barbagallo T , Prestinari F , Ciocca O , Vassallo C , Borroni G
Published: June 2004

Eosinophilic pustular folliculitis (EPF) is an unusual disease, first described in adult East Asians in 1970 by Ofuji. It is characterized by follicular papules and pustules tending to coalesce and form plaques involving the trunk, face and extremities. In recent years, it has been often associated with human immunodeficiency virus (HIV) infection or with immunosuppressed and/or oncohaematological patients. EPF has been described in immunocompetent adult caucasian patients only occasionally. The diagnosis requires clinical and microbiological features such as sterile folliculitis and histopathological findings characterized by folliculitis and perifolliculitis with eosinophilic infiltrate. We describe an HIV seronegative caucasian male with EPF, allergic to non‐steroidal anti‐inflammatory drugs and indomethacin, treated with oral doxicycline. The treatment led to the complete remission of the lesions within 2 months.
Evidence for diversity within Propionibacterium acnes: a comparison of the T‐cell stimulatory activity of isolates from inflammatory acne, endocarditis and the laboratory
Authors: Jappe U , Boit R , Farrar MD , Ingham E , Sandoe J , Holland KT
Published: June 2004

Background  Propionibacterium acnes is primarily associated with the pathogenesis of acne vulgaris but reports are increasing in number implicating P. acnes in other diseases such as abscess formation, meningitis and endocarditis. The pathogenicity of P. acnes is thought to be partly due to the interaction of the bacterium with the immune system. Historically, investigations have focused on humoral and cell‐mediated immune responses to P. acnes antigens without attention to the possibility that different antigens may be expressed by different isolates.

Objective  Investigations were performed to determine whether there were differences between a laboratory strain of P. acnes (P‐37) and fresh clinical isolates in their ability to stimulate naive and adult lymphocytes.

Material and methods  The fresh isolates were collected from a patient with inflammatory acne and a patient with P. acnes‐induced prosthetic valve endocarditis. The lymphocyte transformation assay was used to detect responses to whole‐cell suspensions of stationary phase P. acnes isolates during 7 days of incubation.

Results  The acne isolate was significantly more stimulatory for cord blood mononuclear cells (CBMNCs) than the laboratory isolate (P. acnes P‐37) at day 4 of incubation. There were no significant differences between the three strains at any other time points. However, the isolate cultivated from inflammatory acne was significantly more stimulatory for peripheral blood mononuclear cells (PBMNCs) from acne donors than the endocarditis isolate or the laboratory strain at most time points. There were no significant differences between the endocarditis strain and the laboratory strain.

Conclusion  It can be hypothesized that in case of P. acnes‐induced endocarditis lymphocyte stimulation is a disadvantage for the microorganism and therefore a lack of lymphocyte stimulation may be relevant to the pathogenesis of endocarditis.
Spontaneously remitting dermatomyositis
Authors: Jordan JR , Paruthi S , Pichardo RO , Challa VR , Donofrio PD , Jorizzo JL
Published: June 2004

Dermatomyositis (DM) is an inflammatory myopathy of skeletal muscle with characteristic cutaneous findings. It is a rare disorder with a bimodal age distribution that affects almost twice as many women as men. One category of DM, normal‐enzyme DM, is characterized by cutaneous changes only at baseline, normal serum muscle enzyme levels and myositis demonstrated by electromyography (EMG) and/or muscle biopsy specimens. Typically, patients with normal‐enzyme DM progress to severe muscle involvement and require systemic corticosteroid therapy. The patient we report has normal‐enzyme DM confirmed by serial serum enzymes, EMG, and skin and muscle biopsies but is unique in that she never experienced progression of muscle weakness although muscle involvement was documented histologically and by EMG. Follow‐up examination after 1 year revealed near‐complete resolution of cutaneous involvement after topical therapy and no evidence of muscle weakness.
The borderline syndrome in psychosomatic dermatology Overview and case report
Authors: Harth W , Mayer K , Linse R
Published: June 2004

The borderline syndrome is one of the most severe disturbances of psychosomatic dermatology. Patients with borderline syndrome are situated ‘on the border’ of psychosis, neurosis and personality disorders. The skin as a borderline organ carries a symbolic role. The clinical picture includes artefactual skin diseases due to self‐mutilation by conscious or unconscious cutting, and rubbing, scratching or para‐artefactual manipulations of pre‐existing dermatoses. Leading symptoms of the borderline syndrome are poor impulse control, emotional instability and poor ego strength with low frustration tolerance and unstable personal relationships. We present the case of a 38‐year‐old female patient with borderline syndrome suffering from para‐artefactual skin diseases of the face and a massive hyperhidrosis of the hands and feet. Within 9 months she was treated in four acute psychiatric hospitals and by 12 psychiatrists and psychotherapists. Early and accurate diagnosis and high‐quality, sophisticated long‐term therapy are necessary.
A unified middle class or two middle classes? A comparison of career strategies and intergenerational mobility strategies between teachers and managers in contemporary Hong Kong1
Authors: Wong Yi‐Lee
Published: June 2004

A debate over whether the ‘service class’ is a unitary middle class or is divided between professionals and managers remains unsettled. This qualitative mobility study seeks to elucidate mobility processes of how professional and managerial groups secure their advantages throughout their working lives and over generations. Using data on contemporary Hong Kong, this paper compares the career strategies and intergenerational mobility strategies of forty teachers and thirty managers. Despite employing somewhat different career strategies, managers do not seem less able than teachers to secure their advantages through their worklives. Using the same intergenerational mobility strategies, teachers and managers exploit economic, cultural, and social resources in a similar way for their children's education. So far as demographic stability over generations is concerned a unified service class is in the making.
Women's and men's careers in British sociology1
Authors: Platt Jennifer
Published: June 2004

The career experience of men and women in British university sociology since 1950 is compared, using published data for the whole group and fresh data from a sample of departments. It is shown that, when like is compared with like, the outcomes for men and for women have not been as different as is often suggested; family reasons were more salient in women's careers, but in the end have not made much difference to their destinations. But the proportions of women recruited have varied over time, and the experience of both sexes has been strongly influenced by historical factors affecting different periods. Within the constraints which those have imposed, individuals have not all made the same choices, and that too has affected the outcomes.
The retreat of multiculturalism in the liberal state: theory and policy1
Authors: Joppke Christian
Published: June 2004

This article discusses a recent retreat of multiculturalism in the liberal state. This retreat has occurred both at the level of theory and policy. With the help of some recent liberal critiques of multiculturalism, the first part maps out some shortcomings of the notion of minority integration through cultural recognition, particularly with respect to immigrants. The second part discusses a retreat from multiculturalism policies in three states that had been prominently committed to them: Australia, the Netherlands, and Britain. This practical retreat of multiculturalism is due to a variety of factors, their importance differing across cases: the chronic lack of public support for multiculturalism policies; inherent deficits and failures of multiculturalism policies, especially in socio‐economic respect; and a new assertiveness of the liberal state to impose liberal principles.
Immunohistochemical localization and expression of fibromodulin in adult rat periodontium and inflamed human gingiva
Authors: Qian H , Xiao Y , Bartold PM
Published: June 2004

Objective:  The aim of this study was to determine the distribution and expression of fibromodulin in adult rat periodontal tissues and inflamed human gingiva.

Materials and methods:  The distribution of fibromodulin in rat molar periodontium and human gingival tissue was studied by immunohistochemistry. The expression of fibromodulin mRNA from human gingival fibroblasts, periodontal ligament fibroblasts and osteoblasts was studied by reverse transcription‐polymerase chain reaction (RT‐PCR). For comparative purposes, the distribution and mRNA expression of collagen types I and III, as well as the two small leucine‐rich proteoglycans decorin and biglycan were also studied.

Results:  In the adult rat periodontium, fibromodulin was distributed in the suprabasal gingival epithelium, gingival and periodontal fibroblasts as well as their surrounding extracellular matrices. Strong expression was noted in the palatal gingival tissues and the interfaces of the periodontal ligament with alveolar bone and cementum. In human gingival tissues, staining of fibromodulin was detected in the connective tissue of inflamed gingiva associated with both gingivitis and periodontitis; whereas, weak staining for this molecule was noted in healthy gingival tissues. The expression of mRNA for fibromodulin was strongest in the cultured osteoblasts. Periodontal ligament fibroblasts showed only a weak level of expression for fibromodulin mRNA.

Conclusions:  Fibromodulin is differentially expressed throughout the periodontium being primarily associated with collagen type I in non‐mineralized sites. In addition fibromodulin showed an upregulation in inflamed gingival tissue.
Effects of near‐infrared irradiation to stellate ganglion in glossodynia
Authors: Nakase M , Okumura K , Tamura T , Kamei T , Kada K , Nakamura S , Inui M , Tagawa T
Published: June 2004

Objective:  This study was designed to assess the effect of stellate ganglion near‐infrared irradiation (SGR) on glossodynia and the mechanism of action.

Study design:  Thirty‐seven patients with glossodynia received SGR once weekly for 4 weeks. The response to treatment was evaluated on the basis of the change in pain intensity, assessed with a visual analogue scale (VAS) before and after 4 weeks of treatment. The temperature and blood flow of the tongue were also measured before and after first SGR. As control, eight healthy subjects were studied.

Results:  Tongue pain as assessed by the VAS decreased in 28 of the 37 patients (75.7%). Mean pain intensity decreased significantly from 5.1 ± 2.2 to 1.9 ± 2.1 (P < 0.05). Tongue blood flow at rest in the patients with glossodynia [7.2 ± 1.6 ml min−1 (100 g)−1] was significantly lower than that in the healthy subjects [7.8 ± 0.23 ml min−1 (100 g)−1]. Five minutes after SGR, the temperature of the tongue rose 1.5 ± 0.21°C, and blood flow increased to 8.5 ± 1.2 ml min−1 (100 g)−1. Tongue blood flow (at rest) after 4 weeks of SGR had increased to 7.7 ± 1.1 ml min−1 (100 g)−1.

Conclusion:  SGR is an effective treatment for glossodynia. The mechanism by which SGR improves symptoms associated with glossodynia is thought to be as follows: SGR inhibits abnormally increased sympathetic activity associated with glossodynia. This is followed by normalization of decreased tongue blood flow, thereby alleviating pain.
Ranula: another HIV/AIDS associated oral lesion in Zimbabwe?
Authors: Chidzonga MM , Rusakaniko S
Published: June 2004

Aim:  To show that sublingual ranula is associated with HIV/AIDS and as such should be considered an HIV/AIDS associated oral lesion in Zimbabwe.

Objectives:  To retrospectively study the prevalence, age and gender distribution, the HIV serostatus of ranula patients and the trend in prevalence of ranula and Kaposi's sarcoma (KS) in patients at the two largest referral Oral and Maxillofacial Surgery specialist centres in Harare, Zimbabwe. To use this information to infer an association between ranula and HIV/AIDS in Zimbabwe.

Design:  Descriptive study with a retrospective and prospective component.

Setting:  Oral and Maxillofacial Surgical clinics at specialist referral hospitals, Harare Central Hospital and Parirenyatwa Government Hospital, Harare, Zimbabwe.

Subjects:  Eighty‐three cases of ranula were studied: 45 cases retrospectively and 38 consecutively. A total of 231 cases of KS were studied retrospectively.

Methods:  Histopathologic records of patients who presented with ranula and KS during the period January 1981 to September 2003 were studied. Gender and age were recorded for each case. Thirty‐eight ranula patients studied consecutively during the period June 1999 to September 2003 were consented for HIV testing.

Results:  There were 83 cases of ranula; 43.4% male and 56.6% female. There were 231 cases of KS, 61.2% male and 38.8% female. Male to female ratio was 1:1.3 for ranula and for KS was 1:0.6. Ranula was predominant in the 0–10 year age group (73.5%) while KS was most common in the 21–40 year age group (76.4%). Ranula and KS both had a marked rise in prevalence from 1992 to 2003. A total of 88.5% of the ranula cases tested HIV positive with 95% in the 0–10 year age group.

Conclusion:  There was a rising prevalence of ranula which mirrors that of KS (an HIV/AIDS associated oral lesion) and that 88.5% of ranula patients were HIV positive with 95% of them in the 0–10 year age group. Sublingual ranula should thus be considered another HIV/AIDS associated lesion in Zimbabwe, especially in children.
A Novel Severe Acute Respiratory Syndrome Coronavirus Protein, U274, Is Transported to the Cell Surface and Undergoes Endocytosis
Authors: Tan Yee-Joo , Teng Eileen , Shen Shuo , Tan Timothy H. P. , Goh Phuay-Yee , Fielding Burtram C. , Ooi Eng-Eong , Tan Hwee-Cheng , Lim Seng Gee , Hong Wanjin
Published: June 2004

The severe acute respiratory syndrome coronavirus (SARS-CoV) genome contains open reading frames (ORFs) that encode for several genes that are homologous to proteins found in all known coronaviruses. These are the replicase gene 1a/1b and the four structural proteins, nucleocapsid (N), spike (S), membrane (M), and envelope (E), and these proteins are expected to be essential for the replication of the virus. In addition, this genome also contains nine other potential ORFs varying in length from 39 to 274 amino acids. The largest among these is the first ORF of the second longest subgenomic RNA, and this protein (termed U274 in the present study) consists of 274 amino acids and contains three putative transmembrane domains. Using antibody specific for the C terminus of U274, we show U274 to be expressed in SARS-CoV-infected Vero E6 cells and, in addition to the full-length protein, two other processed forms were also detected. By indirect immunofluorescence, U274 was localized to the perinuclear region, as well as to the plasma membrane, in both transfected and infected cells. Using an N terminus myc-tagged U274, the topology of U274 and its expression on the cell surface were confirmed. Deletion of a cytoplasmic domain of U274, which contains Yxxφ and diacidic motifs, abolished its transport to the cell surface. In addition, U274 expressed on the cell surface can internalize antibodies from the culture medium into the cells. Coimmunoprecipitation experiments also showed that U274 could interact specifically with the M, E, and S structural proteins, as well as with U122, another protein that is unique to SARS-CoV.
Modulation of Env Content in Virions of Simian Immunodeficiency Virus: Correlation with Cell Surface Expression and Virion Infectivity
Authors: Yuste Eloísa , Reeves Jacqueline D. , Doms Robert W. , Desrosiers Ronald C.
Published: June 2004

Specific mutations were created in the cytoplasmic domain of the gp41 transmembrane protein of simian immunodeficiency virus strain 239 (SIV239). The resultant strains included a mutant in which Env residue 767 was changed to a stop codon, a double mutant in which positions 738 and 739 were changed to stop codons, another mutant in which a prominent endocytosis motif was changed from YRPV to GRPV by the substitution of tyrosine 721, and a final combination mutant bearing Q738stop, Q739stop, and Y721G mutations. The effects of these mutations on cell surface expression, on Env incorporation into virions, and on viral infectivity were examined. The molar ratio of Gag to gp120 of 54:1 that we report here for SIV239 virions agrees very well with the ratio of 60:1 reported previously by Chertova et al. (E. Chertova, J. W. Bess, Jr., B. J. Crise, R. C. Sowder II, T. M. Schaden, J. M. Hilburn, J. A. Hoxie, R. E. Benveniste, J. D. Lifson, L. E. Henderson, and L. O. Arthur, J. Virol.76:5315-5325, 2002), although they were determined by very different methodologies. Assuming 1,200 to 2,500 Gag molecules per virion, this corresponds to 7 to 16 Env trimers per SIV239 virion particle. Although all of the mutations increased Env levels in virions, E767stop had the most dramatic effect, increasing the Env content per virion 25- to 50-fold. Increased levels of Env content in virions correlated strictly with higher levels of Env expression on the cell surface. The increased Env content with the E767stop mutation also correlated with an increased infectivity, but the degree of change was not proportional: the 25- to 50-fold increase in Env content only increased infectivity 2- to 3-fold. All of the mutants replicated efficiently in the CEMx174 and Rh221-89 cell lines. Although some of these findings have been reported previously, our findings show that the effects of the cytoplasmic domain of gp41 on the Env content in virions can be dramatic, that the Env content in virions correlates strictly with the levels of cell surface expression, and that the Env content in virions can determine infectivity; furthermore, our results define a particular change with the most dramatic effects.
Transduction Profiles of Recombinant Adeno-Associated Virus Vectors Derived from Serotypes 2 and 5 in the Nigrostriatal System of Rats
Authors: Paterna Jean-Charles , Feldon Joram , Büeler Hansruedi
Published: June 2004

We compared the transduction efficiencies and tropisms of titer-matched recombinant adeno-associated viruses (rAAV) derived from serotypes 2 and 5 (rAAV-2 and rAAV-5, respectively) within the rat nigrostriatal system. The two serotypes (expressing enhanced green fluorescent protein [EGFP]) were delivered by stereotaxic surgery into the same animals but different hemispheres of the striatum (STR), the substantia nigra (SN), or the medial forebrain bundle (MFB). While both serotypes transduced neurons effectively within the STR, rAAV-5 resulted in a much larger EGFP-expressing area than did rAAV-2. However, neurons transduced with rAAV-2 vectors expressed higher levels of EGFP. Consistent with this result, EGFP-positive projections emanating from transduced striatal neurons covered a larger area of the SN pars reticulata (SNr) after striatal delivery of rAAV-5, but EGFP levels in fibers of the SNr were higher after striatal injection of rAAV-2. We also compared the potentials of the two vectors for retrograde transduction and found that striatal delivery of rAAV-5 resulted in significantly more transduced dopaminergic cell bodies within the SN pars compacta and ventral tegmental area. Similarly, EGFP-transduced striatal neurons were detected only after nigral delivery of rAAV-5. Furthermore, we demonstrate that after striatal AAV-5 vector delivery, the transduction profiles were stable for as long as 9 months. Finally, although we did not target the hippocampus directly, efficient and widespread transduction of hippocampal neurons was observed after delivery of rAAV-5, but not rAAV-2, into the MFB.
RBP-J (CSL) Is Essential for Activation of the K14/vGPCR Promoter of Kaposi's Sarcoma-Associated Herpesvirus by the Lytic Switch Protein RTA
Authors: Liang Yuying , Ganem Don
Published: June 2004

The Kaposi's sarcoma-associated herpesvirus (KSHV) gene product virally encoded G protein-coupled receptor (vGPCR) is a homolog of cellular GPCRs and has been proposed to play important roles in KSHV-induced angiogenesis. The most abundant vGPCR-containing transcripts are K14/vGPCR bicistronic RNAs that are strongly induced during lytic reactivation. Here we show that the promoter governing this transcript is strongly responsive to activation by the viral lytic switch protein RTA. By deletion mapping and scanning mutation analyses, we have identified three putative RTA response elements (A, B, and C) in this promoter. However, none of these sites appear to directly bind RTA in electrophoretic mobility shift assays (EMSA). Site C corresponds to a canonical binding site for RBP-J, a sequence-specific transcriptional repressor that is normally the target of Notch signaling. RBP-J can bind RTA and recruit it to its cognate recognition site; when this happens, the activation function of RTA can relieve RBP-J-mediated repression and upregulate expression of the targeted gene. EMSA studies reveal that both sites A and C can bind to RBP-J; sequence inspection reveals that site A is a novel functional variant of known RBP-J recognition sites. (Site B corresponds to an as-yet-unknown host DNA-binding protein.) The importance of sites A and C in vivo is underscored by the observation that K14/vGPCR promoter function is dramatically inhibited in cells genetically deficient in RBP-J. The regulation of K14/vGPCR transcripts by RBP-J raises the possibility that other modulators of Notch signaling might be able to induce expression of this RNA outside the context of lytic KSHV replication.
Characterization of Infectious Retroviral Pseudotype Particles Bearing Hepatitis C Virus Glycoproteins
Authors: Flint Mike , Logvinoff Carine , Rice Charles M. , McKeating Jane A.
Published: June 2004

The recent development of infectious retroviral pseudotypes bearing hepatitis C virus (HCV) glycoproteins represents an opportunity to study the functionally active form of the HCV E1 and E2 glycoproteins. In the culture supernatant of cells producing HCV retroviral pseudotypes, the majority of E2 was not associated with infectious particles and failed to sediment on sucrose gradients. The E2 that was incorporated into infectious particles appeared as a triplet of diffuse bands at 60, 70, and 90 kDa. Similarly, three major forms of E1 were incorporated into the pseudotype particles, migrating at 33, 31, and 25 kDa. Endoglycosidase H (endo-H) treatment of particles demonstrated that the incorporated E1 was partially or completely sensitive to digestion. In contrast, the majority of the incorporated E2 was endo-H resistant. Purified pseudotype particles were found to contain both disulfide-linked aggregates and nonaggregated E1 and E2. HCV pseudotypes generated from cells expressing E1E2p7 showed similar heterogeneity in the incorporated glycoproteins and were of comparable infectivity to those generated by expression of E1E2. Our results demonstrate the heterogenous nature of E1 and E2 incorporated into retroviral pseudotypes and highlight the difficulty in identifying forms of the HCV glycoproteins that initiate infection.
The Human Immunodeficiency Virus Type 1 Promoter Contains a CATA Box Instead of a TATA Box for Optimal Transcription and Replication
Authors: van Opijnen Tim , Kamoschinski Joost , Jeeninga Rienk E. , Berkhout Ben
Published: June 2004

The human immunodeficiency virus type 1 (HIV-1) transcriptional promoter contains a single polymorphism in the TATA box. Most subtypes contain the sequence TATAAGC, but subtype E and some recombinant AG strains have the sequence TA

A

AAGC. Based on mutagenesis studies of cellular RNA polymerase II (pol II) promoters, it has been proposed that the subtype E TATA box is nonfunctional due to the T-to-A substitution at the critical position 3. By means of transcription and virus replication assays, we demonstrate that the true TATA box motif within the viral long terminal repeat (LTR) promoter starts two nucleotides further upstream. Because of this realignment, subtype E has the sequence

CA

TA

A

AA and all other subtypes have the sequence

CA

TATAA. The polymorphism therefore has shifted from position 3 to position 5 and is no longer incompatible with efficient transcription according to rules determined for cellular pol II promoters. In addition, through sensitive competition experiments, we demonstrate that the CATA box of subtypes B and E can be improved for replication by the mutations 1T and 5T, respectively. The fact that the fitness of both subtype LTRs can be increased by specific point mutations in the CATA box suggests that the transcriptional promoter of HIV-1 is fine-tuned towards a suboptimal level of replication. However, this replication rate may be optimal in the in vivo context of an infected individual.
Murine Cytomegalovirus with a Transposon Insertional Mutation at Open Reading Frame m155 Is Deficient in Growth and Virulence in Mice
Authors: Abenes Gerardo , Chan Karen , Lee Manfred , Haghjoo Erik , Zhu Jiaming , Zhou Tianhong , Zhan Xiaoyan , Liu Fenyong
Published: June 2004

A pool of murine cytomegalovirus (MCMV) mutants was previously generated by using a Tn
3
-based transposon mutagenesis approach (X. Zhan, M. Lee, J. Xiao, and F. Liu, J. Virol. 74:7411-7421, 2000). In this study, one of the MCMV mutants, Rvm155, which contained the transposon insertion in open reading frame m155, was characterized in vitro for its replication in tissue culture and in vivo for its growth and virulence in immunodeficient SCID mice. Compared to the wild-type strain and a rescued virus that restored the m155 region, the mutant is significantly deficient in growth in many organs of the infected animals. At 21 days postinfection the titers of Rvm155 in the salivary glands, lungs, spleens, livers, and kidneys of the intraperitoneally infected SCID mice were lower than the titers of the wild-type virus and the rescued virus by 50-, 1,000-, 500-, 100-, and 500-fold, respectively. Moreover, the viral mutant was attenuated in killing the SCID mice, as none of the SCID mice that were intraperitoneally infected with Rvm155 died until 38 days postinfection while all the animals infected with the wild-type and rescued viruses died at 27 days postinfection. Our results provide the first direct evidence that a disruption of m155 expression leads to attenuation of viral virulence and growth in animals. Moreover, these results suggest that m155 is a viral determinant for optimal MCMV growth and virulence in vivo.
Immunization with Hepatitis C Virus-Like Particles Induces Humoral and Cellular Immune Responses in Nonhuman Primates
Authors: Jeong Sook-Hyang , Qiao Ming , Nascimbeni Michelina , Hu Zongyi , Rehermann Barbara , Murthy Krishna , Liang T. Jake
Published: June 2004

We have previously reported the production of hepatitis C virus-like particles (HCV-LP) using a recombinant baculovirus containing the cDNA of the HCV structural proteins (core, E1, and E2). These particles resemble the putative HCV virions and are capable of inducing strong and broad humoral and cellular immune responses in mice. Here we present evidence on the immunogenicity of HCV-LP and the effects of novel adjuvant systems in a nonhuman primate model, the baboon. Three groups of four baboons were immunized with HCV-LP, HCV-LP and adjuvant AS01B (monophosphoryl lipid A and QS21), or HCV-LP and the combination of AS01B and CpG oligodeoxynucleotides 10105. After four immunizations over an 8-month period, all animals developed HCV-specific humoral and cellular immune responses including antibodies to HCV structural proteins and gamma interferon
+
(IFN-γ
+
)CD4
+
and IFN-γ
+
CD8
+
T-cell responses. The immunogenicity of HCV-LP was only marginally enhanced by the use of adjuvants. The overall HCV-specific immune responses were broad and long lasting. Our results suggest that HCV-LP is a potent immunogen to induce HCV-specific humoral and cellular immune responses in primates and may be a promising approach to develop novel preventive and therapeutic modalities.
Production of High-Titer Epstein-Barr Virus Recombinants Derived from Akata Cells by Using a Bacterial Artificial Chromosome System
Authors: Kanda Teru , Yajima Misako , Ahsan Nazmul , Tanaka Mika , Takada Kenzo
Published: June 2004

An Epstein-Barr virus (EBV) genome in Burkitt's lymphoma-derived cell line Akata was cloned into a bacterial artificial chromosome (BAC) vector. The BAC clone, designated AK-BAC, was rapidly and precisely modified by means of efficient homologous recombination in
Escherichia coli
. This system was used to produce recombinant EBVs with transgenes. An expression cassette of green fluorescent protein (GFP) was inserted into AK-BAC, and the resultant BAC clone, AK-BAC-GFP, was transfected into Akata cells. We found that transfected BAC plasmids efficiently formed episomes in EBV-positive Akata cells. Mixtures of wild-type and AK-BAC-GFP viruses were then produced and used to infect EBV-negative Akata cells. We obtained cell clones that harbored only AK-BAC-GFP but no wild-type episome. These cell clones produced infectious viruses after stimulating virus production, and the recombinant viruses of AK-BAC-GFP efficiently immortalized primary B lymphocytes. We further revised the method so that any kind of cDNA could be rapidly inserted into the unique I-PpoI site that had been artificially introduced into AK-BAC. The AK-BAC system will have a broad range of applications, such as genetic analyses of various viral gene products and development of viral vectors for human gene therapy.