Kaposi's sarcoma-associated herpesvirus open reading frame 50/Rta protein activates the entire viral lytic cycle in the HH-B2 primary effusion lymphoma cell line.

Rta, the gene product of Kaposi's sarcoma-associated herpesvirus (KSHV) encoded mainly in open reading frame 50 (ORF50), is capable of activating expression of viral lytic cycle genes. What was not demonstrated in previous studies was whether KSHV Rta was competent to initiate the entire viral lytic life cycle including lytic viral DNA replication, late-gene expression with appropriate kinetics, and virus release. In HH-B2, a newly established primary effusion lymphoma (PEL) cell line, KSHV ORF50 behaved as an immediate-early gene and autostimulated its own expression. Expression of late genes, ORF65, and K8.1 induced by KSHV Rta was eliminated by phosphonoacetic acid, an inhibitor of viral DNA polymerase. Transfection of KSHV Rta increased the production of encapsidated DNase-resistant viral DNA from HH-B2 cells. Thus, introduction of an ORF50 expression plasmid is sufficient to drive the lytic cycle to completion in cultured PEL cells.

Relating speech and swallow function to dropout in a longitudinal study of head and neck cancer.

The relation between functional outcome and dropout from a 12-month follow-up period was examined in a longitudinal study whose objective was to define and quantify the functional effects of oral surgical resection and reconstruction on speech and swallowing in patients with head and neck cancer. In a group of 150 patients recruited to a surgical study in the Cancer Control Science Program in Head and Neck Cancer Rehabilitation, dropout from all causes and dropout from specific causes (medical, patient, and administrative specific) were assessed in relation to longitudinal speech and swallow function. In univariate analysis, better speech articulation was associated with decreased risk of dropout from all causes and from medical-specific causes. Better swallow performance was associated with decreased risk of medical-specific dropout. Multivariate analysis revealed the following: (1) only articulation function was associated with dropout from all causes; (2) the association of speech articulation function with medical dropout was diminished after adjusting for advanced age and surgical resection variables; (3) the association of speech articulation function became significant for patient-specific dropout after adjusting for advanced age and surgical resection variables and indicated that better function decreased the risk of this type of dropout; and (4) swallowing function was not related to dropout. Patients treated for oral or oropharyngeal cancer who have poorer speech outcomes are more likely to drop out from a longitudinal study. Basing study results on only patients who complete a longitudinal study will understate the level of dysfunction experienced.

Factors related to dropout in a study of head and neck cancer patients after surgery.

The extent and nature of dropout was assessed in a longitudinal study whose objective was to define and quantify the functional effects of oral surgical resection and reconstruction on speech and swallowing function in patients with head and neck cancer. Of 150 patients who were enrolled to be followed up with speech and swallow assessments for 1 year after surgery, 113 (75%) dropped out and 37 (25%) returned to complete the study at the final 12-month evaluation point. In general, those completing the study had a smaller resection than the patients who dropped out before the 12-month evaluation. Fifty percent of the dropout was accounted for by medical reasons, 23% by administrative reasons, and 27% by patient-specific reasons (i.e., reasons known only to the patient). Analysis of the dropout categories revealed that higher cancer stage, larger volume of resection, and having a flap surgical closure versus a primary closure or skin graft increased a patient's chance of dropping out. A larger volume of resection was also related to an increased chance of being a patient-specific dropout. Patients who reported no or low alcohol usage had a greater chance of completing follow-up than being a patient-specific dropout.

Functional results of primary closure vs flaps in oropharyngeal reconstruction: a prospective study of speech and swallowing.

BACKGROUND: The preservation of speech and swallowing function is the primary goal when reconstructing soft tissue defects in the oral cavity or oropharynx. The type of reconstructive procedure used should be based on outcome data examining speech and swallowing function; yet, there is a paucity of such information. OBJECTIVES: To present the results of a multi-institutional prospective study of speech and swallowing function before and after soft tissue reconstruction of the oral cavity and oropharynx, and to compare 3 methods of reconstruction with respect to speech and swallowing function: primary closure, distal myocutaneous flap, and microvascular free flap. DESIGN: Prospective case-comparison study. SETTING: Four leading head and neck cancer institutions. PATIENTS: The patients were selected from a database of 284 patients treated at the different institutions. The patients were matched for the location of the oral cavity or oropharyngeal defect and the percentage of oral tongue and tongue base resection. Those patients who had previous speech and swallowing deficits and patients in whom postoperative fistulas or wound infections developed were excluded from the study. METHODS: The patients underwent speech and swallowing evaluation preoperatively and 3 months after healing. This evaluation included videofluoroscopic studies of swallowing and tests of speech intelligibility and sentence articulation. Videofluoroscopy provided measures of swallowing efficiency and bolus movement. Liquid and paste consistencies were used in evaluating swallowing function. MAIN OUTCOME MEASURE: The functional results of the reconstruction. RESULTS: Patients who had primary closure were more efficient at swallowing liquids, had less pharyngeal residue, a longer oral transit time with paste, and higher conversational intelligibility than patients who underwent reconstruction with a distal flap. Compared with patients who underwent reconstruction with a free flap, those who had primary closure had more efficient swallowing of liquids, less pharyngeal residue, and shorter pharyngeal delay times with paste. No difference in the speech and swallowing function existed between patients treated with distal myocutaneous flaps and those treated with microvascular free flaps. CONCLUSION: Contrary to the current theory of oral and oropharyngeal reconstruction, we found that the use of primary closure resulted in equal or better function than the use of flap reconstruction in patients with a comparable locus of resection and percentage of oral tongue and tongue base resection.

Surgical variables affecting speech in treated patients with oral and oropharyngeal cancer.

Postoperative speech function may be influenced by a number of treatment variables. The objective of this study was to examine the relationships among various treatment factors to determine the impact of these measures on speech function. Speech function was tested prospectively in 142 patients with surgically treated oral and oropharyngeal cancer 3 months after treatment. Each patient's speech was recorded during a 6- to 7-minute conversation and while performing a standard articulation test, producing speech outcome measures of percent correct consonant phonemes and percent conversational understandability. Correlational analyses were used to determine the relationships among the speech outcome measures and 14 treatment parameters. Speech function was mildly to moderately negatively correlated with most surgical resection variables, indicating that larger amounts of tissue resected were associated with worse speech function. Overall measures of conversational understandability and percent correct consonant phonemes were related to extent of oral tongue resection, floor of mouth resection, soft palate resection, and total volume of tissue resected. These relationships varied depending on the method of surgical closure. Method of surgical reconstruction had a profound impact on postoperative speech function 3 months after treatment and was an important factor in determining how oral tongue resection influenced articulation and intelligibility. The combination of closure type, percent oral tongue resected, and percent soft palate resected had the strongest relationship with overall speech function for patients with surgically treated oral and oropharyngeal cancer 3 months after treatment.

Medullary carcinoma of the thyroid: prognostic factors and treatment recommendations.

BACKGROUND: Because medullary thyroid carcinoma accounts for only 7% of all thyroid malignancies, data to support treatment strategies are scarce. METHODS: We retrospectively reviewed treatment and outcome in 34 patients with MTC treated at Roswell Park between 1961 and 1995. Univariate analysis was performed using the variables age, sex, tumor size, N stage, and M stage. RESULTS: Median survival was 4.7 years, with 51% and 32% of patients alive at 5 and 15 years, respectively. Nodal metastases were seen in 76% and distant metastases in 67% of all patients. More than 60% of the patients with nodal metastases survived longer than 10 years. Once diagnosed with distant metastases, 90% of the patients died within 5 years. Local failure rate with lobectomy was 44%, compared to 10% after total thyroidectomy (P < .02). Age, extrathyroid extension, and M stage portend a poor outcome. Nodal status had no statistically significant impact on survival. CONCLUSION: Survival with tumors confined to the thyroid gland is independent of nodal status. Long-term survival in patients with distant metastases is rare. This study underscores the role of total thyroidectomy in the initial treatment and the need to develop effective adjuvant therapy for MTC.

Surgery as a single modality therapy for squamous cell carcinoma of the oral tongue.

PURPOSE: The treatment of squamous cell cancer of the oral tongue remains a challenging clinical problem. The efficacy of primary treatment with surgery versus radiation therapy for early stage disease and an adequate treatment paradigm for the clinically negative neck continues to be the subject of clinical debate. We have reviewed our experience in the treatment of oral tongue cancer with surgery as a single definitive treatment modality. PATIENTS AND METHODS: From 1971 to 1993, 79 patients with squamous cell carcinoma of the oral tongue were treated with surgery alone at Roswell Park Cancer Institute. RESULTS: Clinically, 69% of the patients presented with stage I/II disease and 31% presented with stage III/IV. Survival by pathological stage I to IV was 89%, 95%, 76%, and 65%, respectively. Surgical therapy ranged from partial to total glossectomy. There were no patients with positive margins. Local recurrence was observed in 15% of patients with close margins (< 1 cm) and 9% of patients with adequate margins (> or = 1 cm). The incidence of pathological node positive (N+) disease was 6%, 36%, 50%, and 67% for T1, T2, T3, and T4 tumors, respectively. Twenty-five percent of patients undergoing elective neck dissection were pathological N+. All pathological confirmed nodal disease was at level I or II. Of the 43 patients with clinical N0 disease, 16% subsequently developed regional recurrence, all of which were surgically salvaged. CONCLUSION: Locoregional control in patients with squamous cell carcinoma of the oral tongue can be achieved with primary surgical therapy. Adequate margins are crucial to local control. Salvage neck dissection may result in long-term survival for patients with regional relapse. Because of the high rate of occult disease (41%), we currently recommend prophylactic treatment of regional lymphatics for primary clinical disease of T2 or greater.

Squamous cell carcinoma of the floor of mouth: a 20-year review.

BACKGROUND: This study retrospectively examines our treatment choices and outcomes with patients diagnosed with squamous cell cancer of the floor of mouth. Because of our division's past strong surgical bias in the treatment of this disease, we have assessed the results of a patient population treated largely by surgical extirpation. This clinical information has been used to draw conclusions and formulate treatment paradigms for patients with floor of mouth cancer. METHODS: Four hundred fifty patients with the diagnosis of squamous cell carcinoma of the oral cavity received their primary treatment at Roswell Park Cancer Center (RPCI) from 1971 to 1991. Ninety-nine had disease originating in the floor of mouth and are the basis of this retrospective review. RESULTS: Forty-three percent of the patients had early-stage disease (stage I or II). Five-year survival for stages I through IV was 95%, 86%, 82%, and 52%, respectively. The incidence of occult cervical metastases for clinical stage I patients was 21%. For clinical stage II patients, the incidence was 62%. Local control of patients treated with surgery alone was 81%. The regional control rate for these patients was 71%. In patients where negative margins were achieved (> or = 5 mm), the local recurrence rate was 13%, regardless of T stage. Eleven percent of the patients underwent a course of postoperative radiotherapy; all had stage IV disease. When compared with advanced-stage patients undergoing surgery alone, there was a significantly improved regional control rate and a trend toward enhanced survival in the patients receiving adjuvant radiotherapy. CONCLUSIONS: There is a significantly high incidence of occult metastatic disease (21%) for T1 lesions or greater in floor of mouth cancer to warrant elective treatment of regional lymphatics. In patients treated with surgery alone with negative margins, the local control rate was 90% versus 62% when the margins were close or positive. Adjunctive radiotherapy showed a statistically significant (p = .005) increased regional control in patients with stage IV disease. Adjunctive radiotherapy is warranted for increased regional control of disease; good local control can be achieved in floor of mouth cancer with surgery alone when negative margins are obtained.

Identification, expression, and immunogenicity of Kaposi's sarcoma-associated herpesvirus-encoded small viral capsid antigen.

We describe a recombinant antigen for use in serologic tests for antibodies to Kaposi's sarcoma (KS)-associated herpesvirus (KSHV). The cDNA for a small viral capsid antigen (sVCA) was identified by immunoscreening of a library prepared from the BC-1 body cavity lymphoma cell line induced into KSHV lytic gene expression by sodium butyrate. The cDNA specified a 170-amino-acid peptide with homology to small viral capsid proteins encoded by the BFRF3 gene of Epstein-Barr virus and the ORF65 gene of herpesvirus saimiri. KSHV sVCA was expressed from a 0.85-kb mRNA present late in lytic KSHV replication in BC-1 cells. This transcript was sensitive to phosphonoacetic acid and phosphonoformic acid, inhibitors of herpesvirus DNA replication. KSHV sVCA expressed in mammalian cells or Escherichia coli or translated in vitro was recognized as an antigen by antisera from KS patients. Rabbit antisera raised to KSHV sVCA expressed in E. coli detected a 22-kDa protein in KSHV-infected human B cells. Overexpressed KSHV sVCA purified from E. coli and used as an antigen in immunoblot screening assay did not cross-react with EBV BFRF3. Antibodies to sVCA were present in 89% of 47 human immunodeficiency virus (HIV)-positive patients with KS, in 20% of 54 HIV-positive patients without KS, but in none of 122 other patients including children born to HIV-seropositive mothers and patients with hemophilia, autoimmune disease, or nasopharyngeal carcinoma. Low-titer antibody was detected in three sera from 28 healthy subjects. Antibodies to recombinant sVCA correlate with KS in high-risk populations. Recombinant sVCA can be used to examine the seroepidemiology of infection with KSHV in the general population.

Selective switch between latency and lytic replication of Kaposi's sarcoma herpesvirus and Epstein-Barr virus in dually infected body cavity lymphoma cells.

The BC-1 cell line, derived from a body cavity-based, B-cell lymphoma, is dually infected with Epstein-Barr virus (EBV) and Kaposi's sarcoma-associated herpesvirus (KSHV). In these studies, the relationships between these two gammaherpesviruses and BC-1 cells were characterized and compared. Single-cell cloning experiments suggested that all BC-1 cells contain both genomes. In more than 98% of cells, both viruses were latent. The two viruses could be differentially induced into their lytic cycles by chemicals. EBV was activated into DNA replication and late-gene expression by the phorbol ester tetradecanoyl phorbol acetate (TPA). KSHV was induced into DNA replication and late-gene expression by n-butyrate. Amplification of both EBV and KSHV DNAs was inhibited by phosphonoacetic acid. Induction of the KSHV lytic cycle by n-butyrate was accompanied by the disappearance of host-cell beta-actin mRNA. Induction of EBV by TPA was not accompanied by such an effect on host-cell gene expression. Induction of the KSHV lytic cycle by n-butyrate was associated with the expression of several novel polypeptides. Recognition of one of these, p40, served as the basis of development of an assay for antibodies to KSHV in the sera of infected patients. BC-1 cells released infectious EBV; however, there was no evidence for the release of encapsidated KSHV genomes by BC-1 cells, even though n-butyrate-treated cells contained numerous intranuclear nucleocapsids. The differential inducibility of these two herpesviruses in the same cell line points to the importance of viral factors in the switch from latency to lytic cycle.

Detection of human serum antibodies to the BFRF3 Epstein-Barr virus capsid component by means of a DNA-binding assay.

A novel assay for antibodies to an immunodominant component of the Epstein-Barr virus (EBV) capsid antigen (VCA) complex was developed by creation of a chimeric protein containing the DNA-binding domain of the yeast GAL4 protein fused to the capsid antigen encoded by the BFRF3 gene of EBV. GAL4-BFRF3 antigen fusion protein bound specifically to a duplex DNA oligonucleotide containing GAL4-binding sites. Antibodies to the antigen were revealed by retardation of the electrophoretic mobility of the DNA-protein complex. Antibodies to the BFRF3 component of VCA became detectable approximately 2 months after onset of infectious mononucleosis. Kinetics of the antibody response to BFRF3 were identical using supershift or immunoblotting assays. Concordance between the DNA-binding assay and the classical indirect immunofluorescence assay for antibody to VCA was 97%. The GAL4 epitope assay is applicable for detection of antibodies to many cloned gene products.

Clinical experience with a microvascular anastomotic device in head and neck reconstruction.

BACKGROUND: Despite numerous refinements in microsurgical technique and instrumentation, the microvascular anastomosis remains one of the most technically sensitive aspects of free-tissue transfer reconstructions. MATERIALS AND METHODS: Concurrent with the development of microsurgical techniques, various anastomotic coupling systems have been introduced in an effort to facilitate the performance and reliability of microvascular anastomoses. The microvascular anastomotic coupling device (MACD) studied here is a high-density, polyethylene ring-stainless steel pin system that has been found to be highly effective in laboratory animal studies. Despite its availability for human clinical use over the last 5 years, reported clinical series remain rare. Our clinical experience with this MACD in 29 head and neck free-tissue transfers is reported herein. RESULTS: Thirty-five of 37 (95%) attempted anastomoses were completed with 100% flap survival with a variety of donor flaps, recipient vessels, and clinical contexts. Two anastomoses were converted to conventional suture technique intraoperatively, and one late postoperative venous thrombosis occurred after fistulization and vessel exposure. CONCLUSIONS: We conclude that the MACD studied here is best suited for the end-to-end anastomosis of soft, pliable, minimally discrepant vessels. Previous radiation therapy does not appear to be a contraindication to its use. Interpositional vein grafts may also be well suited to anastomosis with the device. When carefully and selectively employed by experienced microvascular surgeons, this MACD can be a safe, fast, and reliable adjunct in head and neck free-tissue transfer reconstructions, greatly facilitating the efficiency and ease of application of these techniques.

Transmissible retrovirus in Epstein-Barr virus-producer B95-8 cells.

Epstein-Barr virus (EBV) released from the B95-8 marmoset cell line has served as a prototype for biologic and biochemical studies of EBV. Here we identify and characterize a retrovirus carried by many cultures of B95-8 cells. The experiments were stimulated by the isolation of a cDNA clone from B95-8 cells in which sequences from the EBV large internal repeat were linked to gag sequences similar to those of squirrel monkey retrovirus, human isolate, SMRV-H. However, among 413 amino acids predicted from the nucleotide sequence of the gag region of the B95-8 SMRV isolate there were 48 amino acid changes that distinguished this virus from SMRV-H originally isolated from a human lymphoid cell line by Oda et al. (1988, Virology 167, 468-476). Nucleic acid and antibody probes were developed for the B95-8 isolate of SMRV. Using such probes, we found that SMRV-B95-8 was readily transmissible, independent of EBV, as an infectious virus to human B and T cell lines. SMRV-B95-8 was highly fusogenic in the presence or absence of EBV. The ultrastructural appearance of the B95-8 retrovirus was characteristic of a type D retrovirus. Cells dually infected with EBV and SMRV-B95-8 did not demonstrate increased levels of lytic EB viral replication. SMRV-B95-8 did not by itself cause lymphocyte immortalization or enhance immortalization by EBV. Thus SMRV-B95-8 does not contribute to the major biologic properties of the B95-8 strain of EBV.

In situ breast carcinoma after treatment during adolescence for thyroid cancer with radioiodine.

We reviewed the courses of patients treated during childhood or adolescence for thyroid cancer to estimate the frequency of, and to identify possible risk factors for, the occurrence of second malignant tumors in this population. We identified all patients treated for thyroid cancer in a cohort of 1,406 pediatric cancer patients who were diagnosed prior to 20 years of age during the period January 1, 1960 through December 31, 1988 and who were treated at Roswell Park Cancer Institute. Twelve patients were treated for thyroid cancer, of whom nine were women. In situ breast carcinoma was diagnosed 25 and 26 years after diagnosis of thyroid cancer in two of four women treated with radioiodine. No new cancers were diagnosed in the five women treated with thyroidectomy only. Two of four women treated for thyroid cancer during adolescence with radioiodine, which is concentrated in the breast as well as other organs, developed in situ breast carcinoma. Review of a large cohort of adolescent female thyroid cancer patients treated with radioiodine is necessary to provide an accurate estimate of their risk of developing breast cancer. These patients must remain under medical surveillance throughout their lifetimes to facilitate prompt diagnosis of and early intervention for new conditions, such as the occurrence of breast cancer.

Anaplastic carcinoma of the thyroid: a 24-year experience.

BACKGROUND: Anaplastic carcinoma of the thyroid gland is a lethal entity; few patients live more than 12 months following diagnosis. We retrospectively reviewed the experience with this entity at our cancer institute and identified a subgroup of patients with complete resection who have a 60% 5-year survival. METHODS: Twenty-one cases of anaplastic carcinoma of the thyroid gland were analyzed retrospectively with respect to prognostic factors influencing survival. This represents 2.7% of 771 cases of thyroid cancer seen at our institution from 1968 to 1992. The median age at presentation was 65.1 years; male/female ratio was 1:1.1; and the most common symptom was a rapidly enlarging neck mass (76%). RESULTS: Estimated 5-year survival was 10% (median: 4.5 months). Tumor size less than 6.0 cm (p = .004) and female gender (p = .02) were significant prognostic factors. Five patients who underwent complete resection had an estimated 5-year survival of 60% (median: 131 months). Four of these patients had postoperative radiotherapy with or without sequential chemotherapy. Two of these patients survived more than 10 years, and a third remains alive without disease at 26 months.

Speech and swallowing function after oral and oropharyngeal resections: one-year follow-up.

BACKGROUND: The purpose of this study was to determine whether the speech and swallowing function of surgically treated oral cancer patients improves between 1 month and 1 year after surgery. METHODS: Speech and swallowing performances were assessed for 28 men and 10 women preoperatively and at 1, 3, 6, and 12 months postoperatively following a standardized protocol. Speech tasks included an audio recording of a brief conversation and of a standard articulation test; swallowing function was examined using videofluoroscopy. Data were also collected on the number and duration of speech/swallowing therapy sessions, as well as the amount and duration of radiotherapy. RESULTS: Statistical analyses revealed that the speech and swallowing function of surgically treated oral and oropharyngeal cancer patients did not improve progressively between 1 and 12 months postsurgery; the level of functioning that these patients demonstrated at the 1- and 3-month posthealing evaluations was characteristic of their status at 1 year after surgery. CONCLUSION: The lack of improvement between 1 and 12 months postsurgery may be related to the relatively small amount of therapy that these patients received during that period. Several outcome variables worsened significantly at the 6-month evaluation; the reversal of function at the 6-month evaluation point could be the effect of postoperative radiotherapy, because irradiated and nonirradiated patients differed in their pattern of recovery on oropharyngeal swallow efficiency and several speech variables.

Surgical variables affecting postoperative swallowing efficiency in oral cancer patients: a pilot study.

This study examined the correlation between swallow function at 3 months postoperatively and surgical variables including volume resected, flap volume, ratio of flap volume to volume resected, percentage of oral tongue, tongue base, and anterior and lateral floor of mouth resected, and whether or not the mandible was preserved in 30 surgically treated oral cancer patients. Swallows of measured amounts of liquid and paste (pudding) materials were examined videofluoroscopically. Nine measures of swallow function were completed for each swallow. A factor analysis of all swallow variables was done for liquid and for paste consistencies to determine whether one measure was statistically representative of all swallow measures. This analysis indicated that oral pharyngeal swallow efficiency (OPSE) represented all measures for both liquid and paste consistencies. Then the correlation between OPSE and surgical variables was defined. Only percentage of oral tongue and percentage of tongue base resected were significantly negatively correlated with OPSE. That is, OPSE decreased for both liquid and paste as percentage of oral tongue or percentage of tongue base resected increased. Results are discussed in terms of diet choices and surgical management.

Transgenic mice and transhybridomas producing chimeric mouse/human anti-human interleukin-2 receptor recombinant antibodies.

Transgenic mice were developed that secreted chimeric mouse/human anti-human interleukin-2 receptor (IL-2R) antibodies (Ab) into their serum. In addition, hybridomas producing the chimeric Ab in tissue culture were generated from the transgenic mice. The presence of the mouse/human immunoglobulin (Ig) transgene did not appear to affect rearrangement of endogenous murine Ig in the hybridomas. Serum levels of the chimeric Ab correlated with transgene copy number. Although many of the transgenic lineages had serum titers of the chimeric Ab comparable to endogenous mouse IgG, there was no apparent correlation with endogenous mouse IgG levels.