Two kinds of antigen suppression in tumor cells revealed by cell fusion.

The Ehrlich ascites tumor, which has been subjected to prolonged selection for growth in allogeneic hosts, possesses powerful mechanisms for the suppression of antigens normally found on the cell surface. It has previously been shown (1, 2) that when cells in which surface antigens are fully expressed are fused with Ehrlich cells, the suppressive mechanisms of the latter continue to operate and the new surface antigens introduced into the hybrid cell by the other partner are also suppressed. In the present paper we describe the properties of hybrids in which one parent cell was the TA3 ascites carcinoma. There are two sublines of this carcinoma which originally arose as a spontaneous mammary carcinoma in a strain A mouse. The TA3/St line has a high concentration of H-2(a) antigens and shows a strain-specific transplantation behavior; the TA3/Ha subline has a drastically reduced antigen concentration and readily transgresses histoincompatibility barriers. The immunoresistant TA3/Ha subline arose spontaneously without having been subjected to any known immunological selection pressure. Hybridization of TA3/Ha cells with normal diploid ACA fibroblasts reestablished full expression of H-2(a) antigens in nine independently derived hybrid clones. Full reestablishment of both D- and K-end components of the H-2(a) complex could be demonstrated. In some hybrid clones the concentration of H-2(a) antigens was found to be comparable to that seen in (A x ACA)F(1) fibroblasts, whereas in others a higher concentration was observed, even exceeding, in some cases, the levels found in the TA3/St line. The H-2(f) complex, contributed by the ACA parent cell, was fully expressed in eight of the nine hybrid clones studied. Antigen suppression thus behaves as a recessive character in the TA3/Ha hybrids, whereas in the Ehrlich hybrids antigen suppression is dominant.

Changed lyotropic liquid crystalline structure due to polymerization of the amphiphilic component.

Optical patterns in polarized light and x-ray reflections in the low-angle region were used to detect a shift from one liquid crystalline structure to another during polymerization. The polymerization took place in a Iotropic liquid crystal of water and sodium undecenoate, with a structure consisting of cylinders in a two-dimensional hexagonal close packing. After polymerization, a lamellar liquid crystalline structure was obtained.

Prognostic value of amplification of c-erb-B2 in bladder carcinoma.

Amplification of c-erb-B2 is examined in patients with transitional cell bladder carcinoma. In a pilot study we found that the amplification correlated with high tumor grade. Tumor grade is a known prognostic factor. Therefore, we next examined the prognostic value of c-erb-B2 amplification in patients with >16 years of clinical follow-up. The gene copy number was determined, using semiquantitative PCR, in archival formalin-fixed tissues. Twenty-three percent (37/163 patients) showed the amplification. The amplification correlated with grade and stage. Moreover, we found that tumor grade (P < 0.001) and c-erb-B2 amplification (P < 0.001) showed prognostic information for survival. Patients with grade 3 tumors and concomitant c-erb-B2 amplification showed the worst prognosis. Multivariate analysis indicates that grade and c-erb-B2 amplification are independent prognostic factors.

Aerobiology in the operating room and its implications for working standards.

Two novel operating room (OR) ventilation concepts, i.e. the upward displacement or thermal convection system and the exponential ultra-clean laminar air flow (LAF) designed to function without extra walls, were evaluated from a bacteriological point of view. The thermal convection system (17 air changes/h) was compared with conventional ventilation (16 air changes/h) with an air inlet at the ceiling and evacuation at floor level. The exponential LAF was compared with the vertical ultra-clean LAF and the horizontal ultra-clean LAF, both with extra side walls. The comparison was made using strictly standardized simulated operations and, except for the horizontal LAF, it was performed in the same OR where the type of ventilation was changed. In the different areas important for surgical asepsis, the thermal system resulted in a twofold to threefold increase in bacterial air and surface counts compared to the conventional system (statistical significance = p < 0.05-0.0001). The bacteriological efficiency of the exponential LAF was equal to the horizontal and vertical LAF units with extra walls in the OR, and all three systems easily fulfilled the criteria for ultra-clean air, i.e. bacteria-carrying particles < 10/m3. In the areas important for surgical asepsis the turbulent ventilation systems yielded highly significant correlation between air and surface contamination (p < 0.02-0.0006). No such correlation existed in the LAF systems.

Silicone/vegetable oil Janus emulsion: topological stability versus interfacial tensions and relative oil volumes.

Several aspects were studied of the formation and destabilization in bulk of silicone/vegetable oil, SO/VO, Janus emulsions, stabilized by Tween 80. In the formation of the emulsions, it was unexpectedly found that the dispersions tended to contain both single and flocculated drops irrespective of the emulsification intensity. Microscopy of the emulsions with no cover glass revealed flocculated drops of a large (200-500 µm) central SO drop with many small VO drops attached. Applying a cover glass did not significantly change the drop size; instead two-oil Janus drops of well-defined contact angle were found. The emulsions showed rapid creaming irrespective of the preparation method, but a few days storage did not significantly change the drop size in the creamed layer, nor was separation of the oils detected. The total interfacial free energy of the Janus drops at equilibrium was compared to the two relevant alternatives; engulfed and separate drops. The Janus drop free energies were found less for all volume ratios of the oils, when the surfactant concentrations in the aqueous phase was sufficient to prevent spreading of VO on SO. Changing the surfactant concentration to bring the interfacial tensions closer to the critical value for spreading gave declining interfacial free energy difference to that of engulfed drops.

Water permeation of reaggregated stratum corneum with model lipids.

Corneocytes were prepared from stratum corneum after extraction of the lipids and then were reaggregated with model lipids to form a membrane. The transport of water through the membrane was found to be similar to earlier published values for reaggregated stratum corneum formed with the indigenous lipids. Similar values were also obtained when only partially saponified free fatty acids were present as lipids. These results support an earlier hypothesis, that the lipid barrier to water penetration of the stratum corneum is determined by the structural organization of the lipids, not by the exact chemical structure of individual species.

Radiotherapy, chemotherapy, and tamoxifen as adjuncts to surgery in early breast cancer: a summary of three randomized trials.

The paper summarizes up-dated results of three randomized adjuvant trials from the Stockholm Breast Cancer Group. The objective of all studies included an evaluation of the role of megavoltage radiation in the primary management of patients with early breast cancer. The first trial was started in 1971 and included 960 pre- and postmenopausal patients with operable disease. The study compared adjuvant radiotherapy with surgery alone. All patients were treated with a modified radical mastectomy. There was a sustained improvement of the recurrence-free survival with radiotherapy (p less than 0.001). Among node positive cases radiation reduced the frequency of both loco-regional recurrence (p less than 0.001) and distant metastasis (p less than 0.01). This observation indicates that distant dissemination in subgroups of patients can originate from uncontrolled local deposits of tumor cells, for instance in the regional lymph nodes. No adverse effect from radiation on long-term survival was observed. The second study was started in 1976 and compared postmastectomy radiation with adjuvant chemotherapy in pre- and postmenopausal high-risk patients. At a mean follow-up of 6 1/2 years there was no significant difference in recurrence-free survival between the two treatments. However, postmenopausal patients fared better with radiotherapy (p less than 0.01). In this subgroup, radiation was more effective than adjuvant chemotherapy in reducing both distant metastases (p less than 0.01) and loco-regional recurrences (p less than 0.001). In the third trial--which only included postmenopausal patients--2 years of adjuvant tamoxifen was compared with no adjuvant endocrine treatment. The number of treatment failures was significantly reduced with tamoxifen (p less than 0.01) but there was no significant overall survival benefit. Subset analysis indicated that tamoxifen improved the recurrence-free survival among patients treated with adjuvant chemotherapy (p less than 0.01) but only to a level close to that achieved with radiotherapy alone. Addition of tamoxifen to radiotherapy failed to further increase the recurrence-free survival.

The promotion of patent airways and inhibition of antigen-induced bronchial obstruction by endogenous nitric oxide.

1. The aim of the present study was to investigate the role of nitric oxide (NO), histamine and leukotrienes in bronchial obstruction. For this, guinea-pigs immunised against ovalbumin were studied under anaesthesia during challenge with antigen or agonists. 2. Challenge with nebulised antigen (0.1-1 mg) elicited dose-dependent increases in insufflation pressure which were abolished by combined administration of histamine and leukotriene antagonists. 3. Challenge with nebulised antigen (0.1-1 mg) also elicited dose-dependent increases in the concentration of endogenous nitric oxide in the exhaled air. After an initial peak, exhaled NO concentrations returned to pre-challenge levels. 4. The increase in insufflation pressure and in exhaled NO caused by ovalbumin challenge was inhibited by combined administration of histamine and leukotriene antagonists. 5. In non-immunised guinea-pigs, challenge of the airways with nebulised histamine (10-1000 nmol) or leukotriene C4 (LTC4, 30-300 pmol) elicited dose-dependent increases in insufflation pressure and in concentrations of endogenous NO in exhaled air. 6. The increase in exhaled NO correlated with the increase in insufflation pressure in response to ovalbumin, histamine and LTC4. An inhibitor of endogenous NO synthesis, N omega-nitro-L-arginine methylester (L-NAME, 30 mg kg-1 i.v.) abolished NO exhalation, and markedly augmented the airway responses to ovalbumin, histamine, or LTC4. 7. The potentiation by L-NAME of the increase in insufflation pressure in response to ovalbumin or histamine was prevented by exogenous NO (20 p.p.m.) in the inhaled air. 8. The results indicate that endogenous NO has an inhibitory effect on bronchial obstruction. Increased NO release during allergen challenge is likely to be due to actions of histamine and leukotrienes.

Chest wall muscle cross talk in canine costal diaphragm electromyogram.

The present paper describes the influence of cross talk from the abdominal and intercostal muscles on the canine diaphragm electromyogram (EMG). The diaphragm EMG was recorded with bipolar surface electrodes placed on the costal portion of the diaphragm (abdominal side), aligned in the fiber direction, and positioned in a region with a relatively low density of motor end plates. The results indicated that cross talk may occur in the diaphragm EMG, especially during conditions of loaded breathing and light general anesthesia. The cross-talk signals showed characteristics that were entirely different from the diaphragm EMG. Although the diaphragm EMG was typical for signals recorded with electrodes aligned in the fiber direction, the cross-talk signals were characteristic of those obtained with electrode pairs not aligned in the direction of the muscle fibers. Alterations in electrode positioning, interelectrode distance, and/or electrode surface area cannot guarantee the elimination of cross-talk signals, whereas spinal anesthesia at a high thoracic level will paralyze the sources of the cross talk and hence eliminate the cross-talk signals. By taking advantage of the differences in EMG signal characteristics for the diaphragm EMG and cross-talk signals, an index that has the capability to detect cross talk was developed.

Transvaginal sonographic findings in ambulatory patients with suspected pelvic inflammatory disease.

OBJECTIVE: To evaluate transvaginal sonographic findings in ambulatory patients with suspected pelvic inflammatory disease (PID). METHODS: We studied 51 outpatients with a mean age of 26.8 years (range 16-52) who had a history of low abdominal pain, negative pregnancy test, and no gynecologic procedures performed during the last month. Endometrial biopsy was used for the histopathologic diagnosis. The presence of plasma cell endometritis was used as the criterion standard for the diagnosis of PID. Sonography was performed before biopsy in a blinded fashion without knowledge of the clinical findings and laboratory results except for the pregnancy test. Repeat pelvic and ultrasound examinations were performed 4 weeks after antimicrobial therapy. RESULTS: Endometrial biopsy revealed plasma cell endometritis in 13 cases (25%). Thickened fluid-filled tubes were seen in 11 of 13 patients (85%) with plasma cell endometritis and in none of those without. Other sonographic findings associated with plasma cell endometritis were polycystic-like ovaries and free pelvic fluid. A sonogram suggestive of PID, ie, thickened fluid-filled tube with or without free pelvic fluid, had a sensitivity of 85% and a specificity of 100% for the diagnosis of plasma cell endometritis. None of the patients with a normal sonogram or simple cyst had plasma cell endometritis. Repeat examination after 4 weeks showed that the sonographic findings had resolved in 60% of the patients who had had histologic evidence of infection. CONCLUSION: Transvaginal sonography can facilitate the outpatient management of patients with suspected PID.

Analysis and presentation. Clinical trials on prostate cancer.

OBJECTIVE: To present guidelines for the analysis and presentation of clinical trials on prostate cancer. METHODS: Textbooks in statistics and oncology were searched for information, as were separate articles on the topic. Previously published advice was fused with own experience. RESULTS: Minimum key points are given for the sections: Introduction, Materials and Methods, Results, Discussion, and Summary. The importance of 1 primary question in any clinical trial is stressed. The value of a detailed presentation of the trial design, the patient population and the inclusion/exclusion criteria, the characterization of the disease, the treatment schedules, and toxicity is underlined. Application of various statistical methods for different endpoints is suggested. Maturity of data, time for publication, and avoidance of publication bias are discussed. Some common pitfalls in the statistical analyses of clinical results are indicated. The impact of prognostic factors, proper staging procedures, and secondary treatments on the interpretation of survival analysis is pointed out. A shift from the (mis-)use of the P value in favor of confidence intervals is strongly encouraged. The use of comparing the survival of responders versus nonresponders is to be abandoned. A few practical hints concerning the presentation are offered. The minimum of data that should be presented in absolute numbers is indicated. Also, the data that should be provided in both graphic and numeric format are exemplified. Examples of essential graphic illustrations are provided. The need for improvements in the design analysis, and presentation of clinical trials is reemphasized. Finally, numerous references are listed. The article is addressed not only to authors and readers of clinical trials, but also to editors of medical journals. CONCLUSION: The suggested guidelines may be useful in the analysis, presentation, and interpretation of clinical trials on prostate cancer. Moreover, compliance with these guidelines may facilitate comparisons with other similar trials and also, the incorporation of single studies into metaanalyses.

Intratumoral measurement and plasma pharmacokinetics of intravenously administered melphalan. Report of a patient with plasmacytoma.

In a human case of plasmacytoma we studied plasma and tumor concentrations of melphalan given intravenously. Intratumoral concentration of melphalan was similar to plasma concentration 60 min after the end of infusion.

Endogenous nitric oxide counteracts antigen-induced bronchoconstriction.

In anesthetized, artificially ventilated guinea pigs immunized against ovalbumin, challenge with aerosolized ovalbumin (0.1 mg) elicited a substantial and sustained increase of insufflation pressure. The inhibitor of endogenous nitric oxide (NO) synthesis, L-NAME (N omega-nitro-L-arginine methylester, 30 mg kg-1 i.v.), markedly augmented the response, the potentiation of which could be prevented by NO (20 p.p.m.) in the inhaled air. The results indicate an inhibitory effect of endogenous NO on antigen-induced bronchoconstriction.

The addition of a mobile ultra-clean exponential laminar airflow screen to conventional operating room ventilation reduces bacterial contamination to operating box levels.

A mobile screen producing ultra-clean exponential laminar airflow (LAF) was investigated as an addition to conventional turbulent/mixing operating room (OR) ventilation (16 air changes/h). The evaluation was performed in a small OR (50 m(3)) during 60 standardized operations for groin hernia including mesh implantation. The additional ventilation was used in 50 of the operations. The LAF passed from the foot-end of the OR table over the instrument and surgical area. Strict hygiene OR procedures including tightly woven and non-woven OR clothing were used. Sedimentation rates were recorded at the level of the patients' chests (N=60) (i.e. the air had passed the surgical team) and in the periphery of the OR. In addition bacterial air contamination was studied above the patients' chests in all 10 operations without the additional LAF and in 12 with the LAF. The screen reduced the mean counts of sedimenting bacteria (cfu/m(2)/h) on the patients' chests from 775 without the screen to 355 (P=0.0003). The screen also reduced the mean air counts of bacteria (cfu/m(3)) above the patients' chests from 27 to 9 (P=0.0001). No significant differences in mean sedimentation rates (cfu/m(2)/h) existed in the periphery of the OR where 628 without and 574 with screen were recorded. During the follow-up period of six months no surgical site infections were detected. In conclusion when the mobile LAF screen was added to conventional OR ventilation the counts of aerobic airborne and sedimenting bacteria-carrying particles downstream of the surgical team were reduced to the levels achieved with complete ultra-clean LAF OR ventilation (operating box).

Inconsistent correlation between aerobic bacterial surface and air counts in operating rooms with ultra clean laminar air flows: proposal of a new bacteriological standard for surface contamination.

The relationship between surface contamination (cfus/m2/h) with particles carrying aerobic bacteria and corresponding air contamination rates (cfus/m3) was evaluated in operating rooms (OR) equipped with ultra clean vertical or horizontal laminar airflow (LAF). For the evaluation we collected data during strictly standardized sham operations using non-woven disposable or cotton clothing. Air contamination in the wound and instrument areas (Casella slit sampler) was related to the surface contamination rate (settle plates) in the same areas and in addition, on the patient chest. Typically, the mean surface counts were 20-70 cfus/m2/h and the air counts 1-2 cfus/m3 in disposable clothing experiments, whilst the use of cotton clothing resulted in higher counts of 100-200 cfus/m2/h (wound P > 0.05, patient P > 0.05, instruments P < 0.01) and 4 cfus/m3 (P < 0.02-0.001). In the vertical LAF, taking both disposable and cotton clothing operations together, the surface and air contamination rates (surface/air ratio SAR) were highly correlated (P = 0.02-0.004) and the ratio varied between 18:1 and 50:1 with a mean for wound air of 36:1. Using only disposable clothing in the vertical LAF, the number of significant correlations was reduced. With cotton clothing experiments in vertical LAF and in the horizontal LAF using disposable clothing, no significant correlation between surface and air contamination was found. The wide variation of SAR values and the inconsistent relationship between surface and air counts indicates that measurement of OR air contamination represents an unhelpful method for assessment of surgical site contamination in LAF units. We propose instead that colony counts on sedimentation plates is a clinically more relevant indicator of bacterial OR contamination in LAF units. In addition to the current bacteriological standard for ultra clean OR air of (< 10 cfus/m3) we suggest a corresponding standard for the surface contamination rate of < 350 cfus/m2/h.

Correlation between surface and air counts of particles carrying aerobic bacteria in operating rooms with turbulent ventilation: an experimental study.

Airborne contamination with bacteria-carrying particles (cfu/m3) and their sedimentation rate (cfu/m2/h) was compared in an operating room (OR) equipped with two turbulent ventilation systems. One was a thermally based system with inlet of cool clean air at the floor level and evacuation of the air at the ceiling by convection (17 air changes/h). The other was a conventional plenum pressure system with air supply at the ceiling and evacuation at the floor level (16 air changes/h). The study was made during rigidly standardised sham operations (N = 20) performed in the same OR by the same six member team wearing non-woven disposable or cotton clothing. Airborne contamination in the wound and instrument areas was related to the surface contamination rate in the same areas and in addition, on the patient chest and in the periphery of the OR. With the exception of the periphery of the OR, the surface and air contamination rates were highly correlated in both ventilation systems (P = 0.02-0.0006, r2 = 0.52-0.79). This was also true particularly when disposable clothing was used while the correlation was weaker in cotton clothing experiments. An equation describing the relation between surface and air counts is given. Typically, the surface counts were numerically 16-fold the air counts, i.e., the number of colonies sedimenting on four 14 cm-diameter agar plates during 1 h will almost equal the number of airborne cfu per m3. We propose, that sedimentation plates represent not only a technically easier method than air sampling but when correctly used, are also the most realistic indicator of airborne bacterial OR contamination in areas critical for surgery.

Mobile zoned/exponential LAF screen: a new concept in ultra-clean air technology for additional operating room ventilation.

A mobile screen (0.5 x 0.4 m) producing ultra-clean exponential LAF (air-flow central zone 0.6 m/s and peripheral zone 0.4 m/s) was investigated as an addition to conventional turbulent/mixing operating room ventilation. The evaluation was performed during strictly standardized sham operations reflecting conditions during major surgery. The study consisted of a pilot experiment designed to give high counts of sedimenting aerobic colony forming units (cfu). In a second main study, recording dust particles, air-borne and sedimenting aerobic cfu, the screen was associated with optimal operating room clothing. In the pilot experiment the use of the screen resulted in a substantial reduction of sedimenting bacteria from 3835-4940 to 0-390 cfu/m(2)/h. In the main study, the use of the additional LAF reduced the surface contamination from 416-329 to 7-78 cfu/m(2)/h up to 1.6 m from the screen (P=0.001-0.0001). Measured in the wound area the screen reduced the air counts of bacteria from 9-14 to 0.2-0.4 cfu/m(3) (P=0.008-0.0001) and a marked reduction of air-borne dust particles was recorded (P=0.007-0.009). In conclusion, the additional mobile LAF screen reduced the counts of aerobic air-borne and sedimenting bacteria-carrying particles as well as dust particles to the levels gained with complete ultra-clean LAF room ventilation. Thus, the screen might prove a valuable addition to operating room ventilation as well as in other areas where asepsis is essential.

Surgical area contamination--comparable bacterial counts using disposable head and mask and helmet aspirator system, but dramatic increase upon omission of head-gear: an experimental study in horizontal laminar air-flow.

The effect of different head coverings on air-borne transmission of bacteria and particles in the surgical area was studied during 30 strictly standardized sham operations performed in a horizontal laminar air flow (LAF) unit. The operating team members wore disposable gowns plus either a non-sterile head covering consisting of a squire type disposable hood and triple laminar face mask, a sterilized helmet aspirator system or no head cover at all. In the wound area both types of head cover resulted in low and comparable air (means of 8 and 4cfu/m(3)) and surface contamination (means of 69 and 126cfu/m(2)/h) rates. Omission of head-gear resulted in a three- to five-fold increase (P > or = 0.01- 0.001), depending on site sampled air contamination rate (mean of 22cfu/m(3)) whereas the bacterial sedimentation rate in the wound area increased about 60-fold ( P > or = 0.0001). A proper head cover minimized the emission of apparently heavy particles that were not removed by the horizontal LAF and contained mainly streptococci, presumably of respiratory tract origin. Dust particle counts revealed no differences between the three experimental situations. No correlation between air and surface contamination rates or between air contamination and air particle counts was found. We conclude that, from a bacteriological point of view, disposable hoods of squire type and face masks are equally as efficient as a helmet aspirator system and both will efficiently contain the substantial emission of bacteria-carrying droplets from the respiratory tract occurring when head cover is omitted. Finally, the use of bacterial air counts to assess surgical site surface contamination in horizontal LAF units must be seriously questioned.

Inefficiency of upward displacement operating theatre ventilation.

A new thermally based ventilation system ('Floormaster') with inlet of cool clean air at floor level, and evacuation at the ceiling of the air warmed by activity in the room (upward displacement ventilation, 17 air changes/h) was compared with a standard positive pressure (plenum) ventilation system with air supply through an inclined perforated screen along one wall at the ceiling and evacuation at floor level (conventional turbulent or mixing system, 16 air changes/h). The study was made during rigidly standardized sham operations (N = 20) performed in the same operating room by a six-member team wearing non-woven disposable or cotton clothing. In general the upward displacement system removed dust particles too small to carry bacteria (0.16-<0.3 microm, 0.001