IB Cell Research Articles

Flow-cytometric DNA analysis of stages IB and IIA cervical carcinoma

The prognostic significance of flow-cytometric DNA analysis was assessed in 375 stages IB and IIA squamous cell carcinoma patients treated with radical hysterectomy and lymphadenectomy at the Mayo Clinic between 1956 and 1985. Paraffin-embedded samples containing at least 20% tumor were dewaxed, rehydrated, stained with propidium iodide, and analyzed. Among 344 assessable samples, 136 (40%) were diploid and 208 (60%) were nondiploid (26 tetraploid, 158 aneuploid, and 24 polyploid). Diploid cases were further subclassified: 25 high proliferative phase (HPP) (S + G 2M > 20%) and 111 low proliferative phase. No significant correlation was noted between DNA diploid patterns and stage, tumor size, grade, or histotype, but HPP diploid tumors had a significantly higher risk of nodal metastasis. With a mean follow-up period of 150 months, 62 patients died of disease. No significant difference was observed in survival rates (SR) between diploid and nondiploid tumors, but the subset of HPP diploid tumors had a prognosis significantly worse than that of any other group ( P < 0.01). Other significant variables included nodal metastases, parametrial extension, age, and clinical stage. While ploidy patterns did not assign additional risk to node-positive lesions, HPP diploid tumors in node-negative patients were associated with a significantly lower SR. Multivariate analyses in node-negative patients demonstrated that stage, histologic subtype, and HPP diploid patterns retained prognostic independence.

酵母表層多糖類の血清学的特異性と抗原決定基と化学構造

We have established an antigenic formula for two serotypes and three subtypes of Saccharomyces cerevisiae by agglutination system with adsorption experiments. This antigenic formulation identifies five distinct antigenic factors or determinants that are found on various serotypes of S. cerevisiae. Serotype Ia and Ib have antigenic factors 1, 10, 18 and 18a and 1, 10, 18 and p, respectively. Serotype Iab has both antigenic factors 18a and p in addition to the common antigenic factors 1, 10 and 18. Serotype II has only the common antigenic factors 1 and 10. 1H-n. m. r. spectrum (H-1 region) of the mannan of serotype Ia, almost free from phosphate, was characterized by the signal at δ5.127 which corresponded to the H-1 of 3-O-substituted α-D-Manp residue adjacent to terminal α (1-3) linkage of mannopentaose side chain, for the specific determinant of antigenic factor 18a. In contrast, phosphodiester mannan from serotype Ib released mannose and α-(1-3)-linked mannobiose by mild acid hydrolysis. The signals at δ5.422-5.461 and 5.284 disappeared, and the signal at δ5.160 was diminished in phosphomonoester mannan treated by mild acid hydrolysis. An immunochemical characterization of the antigenic factor p using agglutination inhibition test between factor p serum and serotype Ib cell ssuggests that the α-(1-3)-linked mannobiosyl-1-phosphate group is a part of the antigenic determinant for the factor p.

Prognostic value of flow cytophotometric DNA content analysis in single treatment stage IB-IIA squamous cell carcinoma of the cervix

DNA content was measured flow cytometrically in archival tissue from 65 single-treatment stage IB and IIA squamous cell carcinomas of the cervix with at least 5 years of clinical followup. Thirty-five cases were treated exclusively by hysterectomy and thirty exclusively by radiation therapy. Tumors were categorized into four groups on the basis of DNA content and cell cycle distribution. DNA content was measured relative to the position of the first resolvable cell peak. G2/M and S-phase fractions were estimated as percentage of cells with DNA contents greater than or equal to relative position 1.70 and percentage of cells with relative positions between 1.20 and 1.70, respectively. The 40 tumors characterized as either aneuploid or nondemonstrably aneuploid with high S-phase fraction estimate had a 5-year recurrence rate significantly higher than that of the 25 tumors categorized as tetraploid or nondemonstrably aneuploid with low S-phase fraction estimate (52 and 4%, respectively; χ 2 = 15.8, P < 0.001). Similar results were found when radiation and surgically treated tumors were considered independently ( χ 2 = 7.95, P < 0.005 and χ 2 = 5.7, P < 0.025, respectively). These data suggest that an increased 5-year recurrence rate is associated with both abnormal DNA content and elevated S-phase fraction in stage IB–IIA squamous cell carcinoma of the cervix, and that this relationship is largely independent of treatment method.

Identification and Characterization of a Functional Receptor for Interferon-γ on a Megakaryocytic Cell Line

We have previously shown that human interferon-gamma (Hu-IFN-gamma) induces platelets to become efficient effector cells, capable of killing young larvae of the parasite Schistosoma mansoni. Recently, binding sites for IFN-gamma on platelets have been characterized. We show here the presence of high-affinity receptors for IFN-gamma on the surface of the human megakaryocytic Dami cell line. Scatchard analysis indicated the presence of about 11,000 binding sites per cell, with a kd of 3 +/- 0.5 x 10(-10) mol/L; the apparent molecular weight of the receptor was 90 Kd. Receptor-bound 125I Hu-recombinant IFN-gamma was rapidly internalized and degraded when the temperature was increased from 4 degrees C to 37 degrees C. The half-life of this receptor was about 7 hours, and pretreatment of cells with IFN-gamma or phorbol myristate acetate had very little effect on the surface receptor number and no detectable effect on IFN-gamma receptor messenger RNA (mRNA) expression. The receptor was functional, because 24 hours of treatment with IFN-gamma led to the increase of HLA class I mRNA expression and to the initiation of HLA class II mRNA expression. These effects were selective because platelet glycoprotein Ib, IIb, or IIIa mRNA expression and cell proliferation were unaffected.

Identification and characterization of a functional receptor for interferon-gamma on a megakaryocytic cell line

We have previously shown that human interferon-gamma (Hu-IFN-gamma) induces platelets to become efficient effector cells, capable of killing young larvae of the parasite Schistosoma mansoni. Recently, binding sites for IFN-gamma on platelets have been characterized. We show here the presence of high-affinity receptors for IFN-gamma on the surface of the human megakaryocytic Dami cell line. Scatchard analysis indicated the presence of about 11,000 binding sites per cell, with a kd of 3 +/- 0.5 x 10(-10) mol/L; the apparent molecular weight of the receptor was 90 Kd. Receptor-bound 125I Hu-recombinant IFN-gamma was rapidly internalized and degraded when the temperature was increased from 4 degrees C to 37 degrees C. The half-life of this receptor was about 7 hours, and pretreatment of cells with IFN-gamma or phorbol myristate acetate had very little effect on the surface receptor number and no detectable effect on IFN-gamma receptor messenger RNA (mRNA) expression. The receptor was functional, because 24 hours of treatment with IFN-gamma led to the increase of HLA class I mRNA expression and to the initiation of HLA class II mRNA expression. These effects were selective because platelet glycoprotein Ib, IIb, or IIIa mRNA expression and cell proliferation were unaffected.

Identification of prognostic factors and risk groups in patients found to have nodal metastasis at the time of radical hysterectomy for early-stage squamous carcinoma of the cervix

In a retrospective study conducted at the University of Alabama at Birmingham, the University of Michigan, and the Mayo Clinic, 185 patients with previously untreated FIGO stage IB and IIA squamous cell carcinoma of the cervix were found to have nodal metastasis at the time of radical hysterectomy and pelvic lymphadenectomy. Of these patients, 103 received adjuvant pelvic irradiation. Cancer recurred in 76 patients; the median time to recurrence was 3.1 years. The prognostic significance of patient age, clinical stage, lesion diameter, number and location of nodal metastases, and use of adjuvant radiation therapy was determined by multivariate analysis. Only patient age ( P = 0.0006), lesion diameter ( P < 0.0001), and number of nodal metastases ( P = 0.0004) were noted to be significant factors in determining overall survival. Rates of recurrence were also related to these factors. Employment of these significant variables led to identification of four risk groups. In general, patients with small cervical lesions (diameter < 1 cm) and no more than two nodes with metastases fell into the low-risk category; those patients with large cervical lesions (diameter > 4 cm) and more than two involved nodes fell into the high-risk category. All other patients were categorized into intermediate-risk groups. Ten-year survival was 92% in the low-risk group ( n = 13), 70% in the low-intermediate-risk group ( n = 66), 56% in the high-intermediate-risk group ( n = 66), and 13% in the high-risk group ( n = 20). This risk group classification identifies subgroups of early-stage cervical carcinoma patients found to have nodal metastasis at the time of radical hysterectomy that warrant appropriately selected adjuvant therapy.

Value of adjuvant whole-pelvis irradiation after Wertheim hysterectomy for early-stage squamous carcinoma of the cervix with pelvic nodal metastasis: A matched-control study

In a retrospective study, 185 patients with previously untreated stage IB or IIA (International Federation of Gynecology and Obstetrics) squamous cell carcinoma of the cervix were found to have pelvic nodal metastasis at the time of Wertheim hysterectomy and bilateral pelvic lymphadenectomy. Of these patients, 103 received adjuvant whole-pelvis irradiation and 82 received no adjuvant therapy. Median dose of pelvic irradiation was 5000 cGy. Among the irradiated patients, in 75% the dose was 5000 cGy or greater. Matching irradiated and nonirradiated patients according to stage, tumor size, and number and location of positive nodes yielded 60 pairs. Mean length of follow-up was 3.9 years for the 60 irradiated patients and 5.8 years for the nonirradiated patients. Kaplan-Meier overall and cancer-specific survival estimates for the two groups were not significantly different ( P > 0.30). During the follow-up period, 21 surgery-only patients and 22 patients treated with adjuvant radiotherapy had recurrence, but adjuvant radiotherapy decreased the proportion of recurrences occurring in the pelvis alone—27% compared with 67% in the surgery-only group ( P = 0.01).

Preoperative adjuvant chemotherapy in the treatment of cervical cancer stage Ib, IIa, and IIb with bulky tumor

Thirty-five previously untreated patients with stage Ib, IIa, and IIb squamous cell carcinoma of uterine cervix with bulky mass (more than 4 cm) were treated with initial chemotherapy of vinblastine, bleomycin, and cis-platinum combined regimen (VBP, one to five courses) and subsequent radical surgery. The effectiveness of the preoperative chemotheapy was evaluated in the surgical specimen. The overall clinical response rate was 89% and included a complete response in 16 (46%) and a partial response in 15 patients (43%). There were no differences in the response rate by age, stage, or the geographic contour of the tumor. The number of chemotherapeutic courses correlated well with the response of the primary tumor ( P = 0.0004) up to three courses. Histologic examination of the resected primary tumor revealed no evidence of disease (Grade IV) in 44% of complete responders, microscopic foci (Grade III) in 38% (6), and macroscopic disease (Grade II) in 18% (3). Of 15 patients with stage IIb, 11 (73%) had a stage-down. Lymphnode metastases after chemotherapy were found in 26% ( 9 35 ) of the patients. All nodal metastases were found among the patients who had a partial response or a stable disease, and none was found in those with a complete response ( P = 0.0029). This preliminary study suggests that initial chemotherapy before surgery is effective in reducing tumor volume or stage of the disease providing better circumstances for surgery, offers selection of high-risk groups of patients requiring additional chemotherapy, and might be able to eliminate effectively diseases in lymphnodes and possibly micrometastases. This regimen is now being evaluated to test its impact on survival.

The prognostic significance of vascular channel involvement and deep stromal penetration in early cervical carcinoma.

Eighty-eight patients with FIGO Stage Ib and IIa squamous cell carcinoma of the cervix underwent radical hysterectomy with pelvic and paraaortic lymphadenectomy. The records and histopathologic material were reviewed to determine the prognostic significance of vascular channel involvement and deep stromal penetration by tumor. Seventy-four patients (84%) were alive and free of disease for more than 2 years and 14 (16%) developed recurrent carcinoma within that time. A positive correlation was found between depth of stromal penetration by tumor and the degree of vascular channel involvement (p less than 0.05). Vascular involvement in itself did not significantly affect nodal status, survival or the rate of recurrence. Depth of penetration was associated with a higher incidence of positive nodes (p less than 0.05). There was a trend towards a lower survival rate and a higher recurrence rate in patients with deep stromal penetration as compared to those with superficial tumors. Microscopic nodal disease increased the rate of recurrence and had an adverse effect on survival. The combination of deep tumor penetration and positive nodes in the same patient was associated with the highest recurrence rate and the lowest survival (p less than 0.05). Nodal status was a more significant prognostic indicator than depth of tumor penetration because patients with deeply penetrating tumors and positive nodes had more than twice the recurrence rate than did patients with deep tumors and negative nodes. Postoperative radiation therapy was beneficial to patients whose tumors demonstrated deep stromal penetration and microscopic metastases to pelvic lymph nodes.

Morphological identification and functional analysis of central neurons innervating the pens retractor muscle of Helix pomatia

1. 1. Cobalt retrograde axonal diffusion and intracellular recordings were used to identify central neurons innervating the penis retractor muscle (PRM) of Helix pomatia. In intact brain-muscle preparations we studied the spontaneous activity of identifiable central neurones and the peripheral effector suspected to be innervated by them simultaneously. 2. 2. Central neurons sending off processes into the penis retractor nerve (PRN) were located in the cerebral ganglia, the right cerebro-pedal connective and the pedal ganglia. Four neurons located on the latero-ventral surface of the right pedal ganglion have been morphologically and electrophysiologically identified revealing the possibility of a routine recognition of individual central neurons from animal to animal. 3. 3. The four identifiable neurons could be classified by virtue of their electrophysiological properties in three types: (1) two irregular bursting cells, (2) one regular beating cell and (3) one silent cell. The firing patterns of the three active cells were found to correspond with rising phase resp. tension maintenance or relaxation of tonic contraction within the PRM. 4. 4. The two irregular bursting cells (IB cells) could be distinguished in IB cell 1 and IB cell 2 by their different burst activities in relation to the muscle behaviour. Spontaneously occurring or' intracellularly stimulated burst activities of IB cell 1 showed a correlation in time with the rising phase of strong contractions within the PRM. The bursting activities of IB cell 1 were not impaired after disconnecting brain and muscle, while the tonic contraction within the PRM was abolished. 5. 5. Spike discharge of the second irregular bursting cell (IB cell 2) accompanied muscle relaxation. Intracellular stimulation of IB cell 2 caused burst activity but failed to produce relaxation from muscle tone. After disconnecting brain and muscle IB cell 2 became silent. 6. 6. Increase in the regular beating rhythm of the third active cell could be observed during tonic muscle activity mainly during the maintained state of muscle tone. After disconnecting brain and muscle this cell fired the whole time in a very regular manner. 7. 7. From these studies it is suggested that IB cell 1 involved in generating strong contractions within the PRM. IB cell 1 seems to exert its effect directly on peripheral nervous structures without the use of centrally located interneurons possibly by a modulatory effect on the efficiency of peripheral motoneurons. I B cell 2 and the regular beating cell appeared to receive direct or indirect afferent input from the peripheral organ. The role of the silent is as yet unresolved.

Viral etiology of age-dependent polioencephalomyelitis in C58 mice

The etiology of immune polioencephalomyelitis (IPE) and the mechanisms of resistance to IPE induction were investigated in C58 mice. IPE was found to be induced by a lipid-solvent-sensitive, filterable replicating agent present in line Ib leukemic cell suspensions. IPE was serially transmitted in immunosuppressed mice with filtered extracts of spleens from diseased animals. The IPE-inducing activity of Ib cell extracts was abolished by chloroform or deoxycholate. Gel filtration of Ib cell extracts showed that the IPE agent has a molecular weight of at least 10(7). Electron microscopy of the active fractions from columns and of spinal cord extracts from mice with IPE revealed a virus-like particle, 40 nm in diameter, which is probably the IPE revealed a virus-like particle, 40 nm in diameter, which is probably the IPE agent. Administration of cyclophosphamide at various times after challenge increased the incidence of IPE in mice, suggesting that IPE is not autoimmune mediated. Immunosuppression resulted in maintenance of high levels of IPE agent in the central nervous system tissue, while immunization resulted in low levels. Moreover, immunized mice produced neutralizing antibodies. These data suggest that antibodies help restrict the amount of IPE agent in the nervous tissue, and that this restriction is required for resistance to IPE induction in C58 mice.

Differential Protective Effects of Immune Lymphoid Cells against Transplanted Line Ib Leukemia and Immune Polioencephalomyelitis

Abstract The capacity of immune cells obtained from the major lymphoid compartments to protect C58 mice from transplanted line Ib leukemia, and from an age-dependent autoimmune CNS disease (immune polioencephalomyelitis = IPE) elicited by immunizing old C58 mice with inactivated Ib cells was quantified. Cells used for comparative adoptive protection tests were harvested from the major lymphoid compartments 14 to 15 days after young C58 mice were immunized with inactivated Ib cell preparations. Regression curves were plotted from survival data and the log10PD50 values were determined. Immune spleen (ISC) and peritoneal cells (IPEC) were significantly more protective against transplanted Ib cells than immune lymph node (ILNC), thymic (ITC), and marrow cells (IMC). In contrast, IPEC and IMC were not protective against IPE and ITC were only marginally protective. ILNC afforded significant protection to transplantable leukemia but were only marginally protective to IPE. When ISC were treated with anti-thy 1.2 serum and complement, protection against transplanted leukemia and IPE was reduced &amp;gt; 99%. When donors of immune lymphoid cells were treated with 12.5 mg of cortisone acetate daily for 2 days before lymphoid cells were harvested, protection against transplanted Ib cells by ISC was reduced by approximately 90% whereas protection against IPE was totally eliminated. Considered together, these results indicate that the protective mechanisms to transplantable leukemia and IPE differ significantly in the same indicator mouse strain.