Signaling reactions induced in human fibroblasts during adhesion to cementum-derived attachment protein.

Cementum-derived attachment protein (CAP) is a Mr 56,000 collagenous protein which promotes the adhesion and spreading of mesenchymal cell types. The CAP promotes the adhesion of osteoblasts and periodontal ligament cells better than gingival fibroblasts, while epithelial cells do not adhere to CAP-coated surfaces. To understand the mechanisms involved in CAP action, we have studied the signal transduction events induced by the CAP in human fibroblasts during cell adhesion. Human gingival fibroblasts were serum starved for 48 h, trypsinized, and added to non-tissue culture plastic plates previously coated with CAP. At various time points, attached cells were examined for induction of signaling reactions. Adherence of cells to plates coated with CAP caused tyrosine phosphorylation of proteins migrating on PAGE with molecular mass of 125-130, 85, 70, and 42-44 kDa. We identified focal adhesion kinase p125FAK and p130Cas as components of the 125-130 kDa protein band; however, p125FAK was the major phosphorylated component. ERK-1 and ERK-2 were detected in the 42-44 kDa protein band, but only the ERK-2, not ERK-1, was phosphorylated. Adhesion to CAP-stimulated mitogen-activated protein kinase (MAPK) activity and induced the expression of c-fos mRNA. Protein-tyrosine phosphorylation and c-fos mRNA expression were not induced in unattached cells, and adhesion was not abolished by the protein tyrosine kinase inhibitor, genestein. MAPK activity and c-fos mRNA expression were not induced in monolayer cultures, indicating that these reactions are induced by adhesion and not necessary for cell adhesion. The kinetics of MAPK activation were different from cells attaching on fibronectin (FN) or polylysine, and c-fos mRNA levels increased only half as much on FN and very little on polylysine. These data demonstrated that CAP and other adhesion molecules present in mineralized tissue matrices induce characteristic signaling events during adhesion, which may play a role in recruitment of specific cell types during wound healing and in mediating their specific biological functions.

Salivary levels of isoniazid and rifampicin in tuberculous patients.

Concentrations of isoniazid and rifampicin were determined in time-matched samples of saliva and serum from 30 tuberculous patients (18 with pulmonary tuberculosis and 12 with intestinal tuberculosis), comprising 18 slow and 12 rapid acetylators of isoniazid, following administration of isoniazid 300 mg and rifampicin 12 mg/kg. The diffusion of isoniazid into saliva was quite rapid and the salivary concentrations were similar to those in serum, suggesting that saliva could be used in place of serum for all pharmacokinetic studies with isoniazid. The salivary concentrations of rifampicin were much lower than those in serum, the mean peak concentrations being 0.9 and 8.5 microgram/ml, respectively. Further, there was evidence of a significant delay in the diffusion of rifampicin from serum to saliva.

Classification of children as slow or rapid acetylators based on concentrations of isoniazid in saliva following oral administration of body-weight and surface-area-related dosages of the drug.

The acetylator phenotype of 180 children aged 3-11 years was determined on the basis of isoniazid concentrations in saliva collected at 5 hours after oral administration of body-weight and surface-area-related dosages of the drug in a syrup form. Isoniazid 2.5 mg/kg was administered on one occasion and 75 mg/m2 surface-area on another, with an interval of 3 days between the occasions. A cross-over design was employed and the sequence was determined by random allocation. The distribution of the concentrations was bimodal with both procedures, indicating the presence of two groups namely, the slow and rapid acetylators. The criterion for a rapid acetylator was a concentration of 0.3 micrograms/ml or less by body-weight-related dosage and 0.4 micrograms/ml or less by that based on surface-area. Based on these criteria, 62% of the children were classified as slow acetylators and 38% as rapid acetylators by body-weight, and 59 and 41%, respectively by surface-area, and the findings were similar in children in the different age-groups. The agreement between the two procedures was 98%.

Induction of rifampicin metabolism during treatment of tuberculous patients with daily and fully intermittent regimens containing the drug.

Self-induction of rifampicin metabolism during daily and intermittent chemotherapy was studied by monitoring the changes in the serum half-life of the drug over a 4-week period in patients with pulmonary tuberculosis. Rifampicin 450 mg was administered to 8 patients who received treatment daily, 7 on thrice-weekly and 7 others on twice-weekly treatment. Serum half-life was computed from concentrations of the drug determined at 3, 4 1/2 and 6 hours after drug administration, on admission and at 1, 2 and 4 weeks after start of treatment. In the daily series, the mean serum half-life decreased from 4.9 hours on admission to 3.6 hours at 1 week (P = 0.02), and treatment beyond this had no further effect. In the thrice-weekly series, maximal induction was observed at the 2nd week, the mean values on admission and at 2 weeks being 5.8 and 3.7 hours, respectively (P less than 0.01). In the twice-weekly series, maximal induction was observed only at the 4th week, the mean values on admission and at 4 weeks being 4.9 and 3.7 hours, respectively (P less than 0.01). Serum activity of gamma glutamyl transferase was not found to be a suitable in vivo marker to monitor induction of the hepatic microsomal enzymes as no significant changes were observed in the activity of this enzyme in any of the 3 series during the 4-week period.

Rifampin-induced release of hydrazine from isoniazid. A possible cause of hepatitis during treatment of tuberculosis with regimens containing isoniazid and rifampin.

The effect of daily administration of rifampin on the direct conversion of isoniazid to isonicotinic acid and hydrazine by isoniazid hydrolase was investigated in 6 slow and 8 rapid acetylators of isoniazid. The proportion of isoniazid metabolized through this direct pathway during the first 6 h was estimated from the ratio of total isonicotinic acid formed to acetylisoniazid in urine after administration of isoniazid or acetylisoniazid. In slow acetylators, this proportion was approximately 3% when isoniazid alone was administered and approximately 6% during the maximal phase of induction caused by the daily administration of rifampin in addition to isoniazid (p less than 0.001); in rapid acetylators, the proportions were considerably less (less than 1 and 2.5%, respectively), suggesting that isoniazid hydrolase was induced by rifampin. The increased formation of hydrazine, a known hepatotoxic agent in animals, could explain the substantially higher frequency of the occurrence of hepatitis in slow than in rapid acetylators among tuberculous patients treated with daily rifampin and isoniazid.

Use of intravesical cisplatin and mitomycin-C for recurrent transitional cell carcinoma of bladder refractory to thiotepa.

Superficial transitional cell carcinoma of the bladder (TCCaB) has a high incidence of recurrence. Intravesical thiotepa is the drug used most often for treatment. Intravesical mitomycin-C also has shown promise. Cisplatin has proven activity in patients with metastatic TCCaB and in experimental bladder cancer models. Patients who have no response to one intravesical agent such as thiotepa may respond to other agent(s) such as mitomycin-C or cisplatin. In this report we have evaluated 10 patients with superficial recurrent TCCaB who showed no response to intravesical thiotepa and were subsequently treated with mitomycin-C or cisplatin. Six patients were treated with cisplatin and 4 with mitomycin-C. The 4 treated with mitomycin-C had complete response or partial response within three months. Of the 6 treated with cisplatin, 4 had complete response within three months, and 2 had no response.

Pelvic lymphadenectomy for carcinoma of prostate.

Pelvic lymphadenectomy was performed in 187 patients as a staging procedure for carcinoma of the prostate. Clinical impression of lymph nodes during dissection appears to be inaccurate in a large number of patients. We therefore believe that if radical surgery is planned for clinically localized tumor, the histologic status of the removed nodes should be confirmed before surgical extirpation of the tumor.

Cystadenocarcinoma of the kidney: a profile of 13 cases.

Cystadenocarcinoma is an uncommon variant of renal cell carcinoma that is cystic in appearance. Our experience with 13 of these tumors is presented. The biological behavior was similar to renal cell carcinoma in general, although a greater proportion presented as stage I lesions. Computerized tomography and cyst aspiration may be helpful to differentiate these tumors from simple cysts.

Prostatic carcinoma: detection and staging using suprapubic US.

Eighteen patients with pathologically proved carcinoma of the prostate were examined with suprapubic ultrasound (US) to assess the capability of the modality in the detection and staging of the disease. Five of the patients were evaluated for recurrent disease. A detailed, retrospective, echo-textural analysis of the prostate and periprostatic structures was performed. The most frequently encountered US findings were recorded and used as criteria in the diagnosis and staging. Most diagnostic of the echo patterns were the presence of a large (greater than 6 mm) focal density within the peripheral zone that was as bright as or less bright than the density of the capsule, a mixed or diffuse pattern, and loss of integrity of the capsule, seminal vesicles, or bladder. Using these criteria, we were able to correctly diagnose 17 of 18 cases, with an accuracy rate of 94%. US was also used to assess the extent of local disease in carcinoma of the prostate. The US staging was compared with the staging by digital rectal examination and by clinicopathologic results. We correctly staged 15 of 18 cases (83%) with US, while the rectal examination helped to stage correctly only 10 of 18 cases (56%). All errors were due to understaging. We find suprapubic US to be very helpful in the diagnosis and staging of prostatic cancer. Since there is some overlap between the findings in benign and malignant disease, we cannot recommend US as an alternative to prostate biopsy. Further experience in detection and staging with prospective studies may be necessary before final conclusions can be made.

Radical prostatectomy with adjuvant radioactive gold for prostatic cancer: a preliminary report.

A total of 71 patients with prostatic carcinoma underwent radical prostatectomy combined with instillation of radioactive gold seeds between January 1, 1977 and July 1, 1982. Gold seeds were implanted intraoperatively into the region of the prostatic vascular pedicles and surrounding fibrolymphatic tissue after removal of the prostate and seminal vesicles. Of the patients with disease confined to the prostate clinically 46.7 per cent had microscopic extraprostatic local tumor extension. We believe that patients with extraprostatic local tumor derive therapeutic benefit from the addition of adjuvant radioactive gold seeds at prostatectomy. Because of the inaccuracy of clinical staging by rectal examination, we have elected to use radioactive gold seeds in all of our patients at prostatectomy. To date, local recurrence has been noted in 5.6 per cent of our patients and systemic progression in 14.1 per cent.

Urinalysis: the physician's responsibility.

Careful examination of a fresh, properly collected urine specimen can indicate the presence of urinary tract disease or uncover unsuspected local or systemic pathology. Urinalysis also can lead to a more complete understanding of a systemic disorder or can assure both the patient and the physician that urinary tract disease is absent, at least within the limitations of the examination. Performing a urinalysis takes no more than 10 minutes, is relatively inexpensive and requires minimal training in proper techniques.

Urinary bladder tumors studied by cystosonography. Part I: Detection.

Findings of excretory urography, the principal screening test for bladder tumors, do not accurately depict these tumors. In an attempt to assess the value of sonography in detecting bladder lesions, we used suprapubic sonography to examine 120 patients who presented with various lower urinary tract symptoms. Commercially available equipment was used. Ultrasound detected 16 of the 17 cases of bladder tumor that were confirmed by cystoscopy. In 14 of these, satisfactory images were obtained. Nontechnical factors that affect detection include size and location of the tumor, bladder distention, obesity of the patient, and the operator's skill. Blood clot, benign prostatic hypertrophy, cystitis, and bladder trabeculae can mimic bladder tumors. Sonography may be helpful as an initial screening test for patients suspected of having bladder tumors; however, further experience is needed before conclusions can be made.

Urinary bladder tumors studied by cystosonography. Part II: Staging.

Correlation between the sonographic staging and the clinical staging as determined by excretory urography, CT, cystoscopy, bimanual examination under anesthesia, and biopsy was possible in 14 of 17 cases of bladder tumor. Ultrasound failed to stage three cases either because the tumor was not seen by sonography (one case) or because of technically suboptimal images (two cases). The cystoscopically obtained biopsy specimens were adequate to make pathologic staging possible in 12 of 14 cases. There was agreement between the sonographic and clinical staging in all 14 cases and with the pathologic staging in all 12 adequately staged cases. The shortcomings of pathologic staging using the cystoscopic biopsy and the criteria of sonographic staging are discussed. Although sonography was demonstrated to be more accurate than conventional methods, further experience is necessary before conclusions can be drawn.

Combination perineal cryosurgery and external radiation therapy for adenocarcinoma of prostate.

Perineal cryosurgical destruction of the prostate has been practiced at the University of Iowa since 1969 for a select group of patients with primary adenocarcinoma of the prostate. We have reviewed this technique in the past and analyzed it with respect to both patient survival and elimination of the local lesion. In this report we have evaluated 14 patients who received combination cryosurgery and external radiation therapy for prostatic adenocarcinoma. Several clinical parameters were examined with emphasis placed on control of the local lesion and obviating the need for urinary diversion or transurethral resections to relieve obstructive symptoms.

The frozen section in lymphadenectomy for carcinoma of the prostate.

From January 1969 through August 1982 we performed 257 pelvic lymph node dissections for staging of carcinoma of the prostate. One or more lymph nodes were submitted for frozen section in 100 cases (39 per cent): 43 (43 per cent) had positive nodes on frozen and permanent sections, while frozen section was negative in 57 (57 per cent). Of the latter 57 cases (16 per cent) had positive nodes on paraffin sectioning of all tissue submitted. Thus, in our series there was a 16 per cent chance of overlooking metastatic disease at the time of pelvic lymph node dissection even with the aid of intraoperative frozen sections.

Catheter-induced cystitis: evaluation by cystosonography.

The bladders of 23 patients with indwelling catheters were examined by ultrasound and cystoscopy. Twelve of the 23 showed changes consistent with bullous cystitis, a catheter-induced reaction of the bladder mucosa that may simulate a bladder tumor on intravenous urography and sonography. The changes seen included a thickened mucosa that was smooth in the early stages, becoming redundant and polypoid in later stages. The mucosa was usually hypoechoic and the lesions were localized on the posterior wall or were diffuse and more severe, depending on the length of catheterization. It is concluded that the sonographic findings in the appropriate clinical setting are suggestive of this entity. Although cystoscopy is not often indicated, cystoscopic findings are diagnostic and biopsy is, therefore, unnecessary.

Urinary tract infection and risk of bladder cancer.

In an epidemiologic study of 2982 bladder carcinoma patients and 5782 population controls from 10 geographic areas of the United States, the role of urinary tract infection and inflammation in the etiology of this neoplasm was evaluated. A history of urinary tract infection significantly elevated the risk of bladder cancer, particularly in individuals who reported three or more infections (relative risk (RR) = 2.0). Significantly increased bladder cancer risk was also found for bladder stones (RR = 1.8), while kidney stones showed no relation. A history of three or more urinary tract infections was strongly related to squamous cell carcinoma in particular (RR = 4.8).