[Benefits of the classical approach in surgery for pulmonary metastases]. / Prednosti klasického operacního prístupu v chirurgické lécbe plicních metastáz.

INTRODUCTION: Distant metastases remain a significant problem in the treatment of malignancies. Surgical management of pulmonary metastases is considered valuable from the oncological view only on condition that R0 resection can be achieved. The whole spectrum of resection procedures can be used, however most commonly, extraanatomic lung resections are employed. It has not been fully evaluated whether the same efficacy can be obtained with thoracoscopic procedures. AIM OF THE STUDY: The aim was to compare the study complication rates with literature data. The secondary aim was to evaluate the benefit of intraoperative lung palpation examination. MATERIAL AND METHODS: The authors present a retrospective study in a group of subjects operated for secondary pulmonary malignancies in the Motol Charles University 2nd Medical Faculty and Faculty Hospital Surgical Clinic, from 2003 to 2007. The authors compared the patient group's morbidity and 30-day mortality rates with literature data. Preoperative CT findings, intraoperative palpation findings and histological examination findings were assessed. RESULTS: Postoperative morbidity of the operated subjects was 16.5%, postoperative 30-day mortality was 0%. The authors compared the preoperative diagnostic data based on CT, the intraoperative findings and histological findings. During the total of 77 surgical procedures, including open and VATS procedures, the authors performed intraoperative palpation examination and detected 60 foci (24.6% out of the total removed foci) previously undetected on CT. All of the foci were of less than 5mm and in 55 cases, the foci were proved metastases. CONCLUSION: The outcome data showing low postoperative morbidity rates and nul 30-day mortality have confirmed that pulmonary metastasectomy is a safe method, a part of the complex oncological management. A surgeon's palpation finding is considered unsubstitutable in the detection of all lung foci and for necessary orientation in order to identify the safety margin in wedge resections. Therefore, the authors prefer the open or videoassissted approach to purely miniinvasive procedures.

CD5 is a potential selecting ligand for B cell surface immunoglobulin framework region sequences.

In rabbits nearly all B lymphocytes express the glycoprotein CD5, in contrast to mice and humans, where only a small proportion of B cells express this molecule (Raman, C., and K.L. Knight. 1992. J. Immunol. 149:3858-3864). CD5+ B cells appear to develop early in ontogeny and be maintained throughout life by self-renewal. The function of CD5 on B cells is still unknown. We showed earlier that "positive" selection occurs during B lymphocyte development in the rabbit appendix. This selection favors B cell expressing surface immunoglobulins with VHa2 structures in the first and third framework regions (Pospisil, R., G.O. Young-Cooper, and R.G. Mage. 1995. Proc. Natl. Acad. Sci. USA. 92:6961-6965). Here we report that F(ab')2 fragments, especially those bearing VHa2 framework region determinants, specifically interact with the B cell-surface glycoprotein CD5. This interaction can be inhibited by anti-CD5 antibodies. Furthermore, immobilized F(ab')2 fragments selectively bind CD5 molecules in appendix cell lysates. Interactions of VH framework region structures with CD5 may affect maintenance and selective expansion of particular B cells and thus contribute to autostimulatory growth of autoimmune or transformed cells.

Free latissimus dorsi muscle flap for chronic bronchopleural fistula.

The author presents experience with the treatment of chronic bronchopleural fistula in a patient who suffered a gunshot injury to his right chest. During the primary assessment the patient underwent resection of two lobes of the right lung, and the patient healed without complications. Four years later the patient presented with bronchopleural fistula with empyema. We had to proceed with right sided pneumonectomy and close up the fistula. The fistula recurred, and the patient underwent repeated operations by chest surgeons without success. Finally, the situation was solved by a contralateral latissimus dorsi muscle free flap. The free-flap was wrapped around the bronchial stump, and the muscle also filled the chest cavity. We discuss the post-surgical course and present the final result.

[Bronchoplastic surgery for non small-cell lung cancer]. / Bronchoplastické operace pro nemalobunecný plicní karcinom.

Bronchoplastic surgery enlarge the spectrum of standardlung resections as a treatment of non-small cell lung career with preserved oncological radicality. The aim of our study is to assess, based on our own experience with 39 patients that underwent this kind of surgery, the results in terms of post-operative morbidity and mortality, to point out the risks of complication linked with this kind of resection and to define the factors suitable to minimize these risks. Very good immediate as well as long-term results place the bronchoplastic surgery to the same level of importance to standard lung resections. In case of the convenient localization of the central tumors, there is an indication for all patients regardless of the respiratory function parameters.

Rabbit appendix: a site of development and selection of the B cell repertoire.

As early as 1963, it was proposed that the rabbit appendix was a homologue of the chicken bursa of Fabricius (ARCHER et al. 1963). The finding that the young rabbit appendix was thymus independent contributed to the concept of central primary lymphoid tissue. Today we know that appendix is a site that generates the high copy number primary repertoire through diversification of rearranged VH genes by gene conversion-like and somatic hypermutation mechanisms. Thus the appendix of young rabbits functions as a mammalian bursal equivalent. In the appendix, newly generated B cells also undergo selection processes involving self and foreign antigens and superantigens. Preferential expansion and survival of B cells in normal and mutant ali rabbits based on FR1 and FR3 expression may involve "superantigen"-like interactions with endogenous and exogenous ligands. One endogenous ligand appears to be CD5. Additional ligands may be produced by gut flora. Further studies in the rabbit model are needed to determine the fates of emigrants from primary GALT, their sites of postulated self-renewal in the periphery, and the nature of secondary diversification in secondary germinal centers where populations of B lymphocyte memory cells may develop. These data may also be helpful in understanding how the repertoire of human B cells is formed and how this repertoire might be manipulated for clinical benefit.

The effect of food on pharmacokinetics and pharmacodynamics of fenoldopam in class III heart failure.

Eighteen patients with New York Heart Association class III congestive heart failure were given single 100 mg oral doses of fenoldopam with food or fasting in a random-order single-blind crossover trial. Before and after each fenoldopam dose, thermodilution cardiac output, right atrial pressure, pulmonary artery pressure, and pulmonary capillary wedge pressure (PCWP) were measured with a balloon-tipped pulmonary artery catheter, and heart rates and blood pressures were recorded with an automated sphygmomanometer. Compared with fasting, bioavailability of fenoldopam was decreased significantly when administered with food: mean peak plasma fenoldopam level decreased from 26.5 (+/- 4.1 SEM) ng/ml to 10.9 (+/- 1.7 SEM) ng/ml (p = 0.0004) and mean area under the concentration-time curve was decreased from 44.7 (+/- 5.8 SEM) ng.hr/ml to 26.8 (+/- 4.1 SEM) ng.hr/ml (p = 0.0001). Fenoldopam administration to fasting patients resulted in decreases in mean arterial pressure, systemic vascular resistance, and PCWP and significant increases in cardiac index without change in heart rate. The maximum changes in mean cardiac index, systemic vascular resistance, and PCWP were greatest 1 hour after oral administration and did not persist beyond 3 hours after administration. In fasting patients, changes in cardiac index were correlated with plasma fenoldopam levels, whereas changes in PCWP and mean arterial pressure did not correlate significantly with the observed fenoldopam level.

Skin response to tuberculin in pig foetuses and germ-free piglets.

Cellular response to intradermally administered PPD (2 TU) was demonstrated in pig foetuses of various ages and in germ-free piglets. CD2+, CD4+, CD8-, 86D-, SLA-D- T lymphocytes were the predominant cells in the skin tuberculin reaction in both foetuses and germ-free animals. Reactive T cells were observed as early as in mid-gestation, whereas SLA-D+ (porcine MHC class II) cells appeared only in older foetuses. Polymorphonuclear leukocytes were never observed in the PPD reaction.

Assessment of hemodynamic tolerance from a 24-hour intravenous infusion of fenoldopam mesylate in congestive heart failure.

To determine the maintenance of pharmacodynamic effects of fenoldopam mesylate, a dopamine-1 agonist, the invasive hemodynamic profiles of 33 patients with New York Heart Association functional class III to IV congestive heart failure were examined. Fenoldopam mesylate was initiated at 0.1 micrograms/kg/min and titrated to a cardiac index greater than or equal to 25% above baseline. Upon achievement of optimal hemodynamics, maintenance infusion was begun (mean dose 0.6 micrograms/kg/min). Fenoldopam mesylate (baseline vs maximal effect) decreased systemic vascular resistance by 37% (p less than 0.001), left ventricular filling pressure by 16% (p less than 0.05) and mean arterial pressure by 11% (p less than 0.05), with an associated augmentation in cardiac index and stroke volume index by 27% (p less than 0.001). Attenuation of hemodynamic effect (maximal effect vs time) was noted in cardiac index (14% p less than 0.001), systemic vascular resistance (13% p less than 0.05) and stroke volume index (13% p less than 0.05). None of the parameters exhibited complete attenuation to baseline values. Fenoldopam mesylate improves cardiac output and lowers systemic vascular resistance with relative attenuation of pharmacodynamic effect during a 24-hour intravenous infusion.

Characterization of antigen-binding protein in earthworms Lumbricus terrestris and Eisenia foetida.

Injection of antigen into the annelid worms Lumbricus terrestris (LT) and Eisenia foetida (EF) results in a marked increase of coelomic fluid protein concentration and the formation of a protein which binds the stimulating antigen (3). In this report we show that the increases in total protein concentration after first and second doses of antigen were higher and were achieved earlier in LT than in EF, while the accumulation of antigen-binding protein in coelomic fluid was similar in both species. Antigen-binding protein isolated by affinity chromatography retained its original binding activity. Its molecular weight in coelomic fluid as well as after isolation was 56 kD when analyzed by SDS-PAGE and immunoblotting. Under reducing conditions, two bands with mol./wt. 31 and 33 kD appeared which did not reveal detectable binding activity. This suggests that the 56 kD binding protein of annelids is composed of two disulphide-linked polypeptide chains both of which participate in antigen-binding site formation.

Continuous monitoring of mixed venous oxygen saturation as an indicator of pharmacologic intervention.

This prospective study evaluated the ability of continuous SvO2 measurements to predict the onset and duration of action of the oral vasodilator, fenoldopam. Eight patients with New York Heart Association functional class 3 CHF received 100 mg fenoldopam in the fasted state. Serial hemodynamic parameters and SvO2 measurements were obtained at baseline and up to eight hours postdose. Although wide interpatient variability was observed, the SvO2-time response curve produced a similar trend as the CI-time response curve. The SvO2 may be a useful drug monitoring parameter in patients with NYHA class 3 CHF. Seriously ill patients may not have a good CI/SvO2 correlation. This is most likely due to an unstable oxygen consumption rate at the tissue level. Therefore, we recommend establishing the relationship between CI and SvO2 prior to its use as a drug monitoring parameter.

CD5 is A potential selecting ligand for B-cell surface immunoglobulin: a possible role in maintenance and selective expansion of normal and malignant B cells.

Although the function of CD5 on B cells is unknown, previous studies suggested that CD5 interaction with V(H) framework regions of surface immunoglobulins (Igs) may contribute to survival and expansion of B cells. Here we used B-chronic lymphocytic leukemia (B-CLL) cells and transformed B-cell lines from normal and B-CLL patients to study CD5-Ig interactions. Immobilized Ig binds and permits isolation of CD5 from lysates of CD5-expressing cell lines. Immunoglobulins or Fab fragments of different V(H) families varied in their effectiveness as inhibitors of anti-CD5 staining of CLL cells, appendix and tonsil tissue sections. Human Ig also binds to purified recombinant CD5. We show here for the first time that the unconventional Ig-CD5 interaction maps to the extracellular CD5-D2 domain whereas conventional epitopes recognized by anti-CD5 antibodies are localized in the D1 domain of CD5. We propose that interactions of VH framework regions with CD5 as a ligand may maintain, select or expand normal, autoimmune or transformed B cells and also contribute to skewing of the normal V(H) repertoire.

Invasive pharmacodynamic characterization of combined ibopamine and calcium blocker therapy for heart failure.

STUDY OBJECTIVE: To determine the acute hemodynamic response of single-dose coadministration of ibopamine plus nifedipine or diltiazem in patients with New York Heart Association functional class (NYHA FC) II-III congestive heart failure. DESIGN: A single-blind, placebo-controlled, two-paired, crossover study. SETTING: Cardiology clinics at two large teaching hospitals. PATIENTS: Eight patients with NYHA FC II-III congestive heart failure who met the inclusion criteria were selected randomly. INTERVENTIONS: All patients underwent right heart catheterization. Day 1 consisted of concomitant calcium channel blocker plus placebo, with cardiac and peripheral hemodynamic recordings from 30 minutes-24 hours. The design was equivalent on day 2, with single-dose administration of ibopamine plus calcium channel blocker. MEASUREMENTS AND MAIN RESULTS: Single-dose nifedipine-diltiazem augmented cardiac output and stroke volume secondary to decreasing systemic vascular resistance. The nifedipine-ibopamine and diltiazem-ibopamine subgroups demonstrated relatively equal hemodynamics, augmenting cardiac index (nifedipine 43%, p < 0.05; diltiazem 40%, p < 0.05 vs baseline) while decreasing systemic vascular resistance (nifedipine 41%, p < 0.05; diltiazem 28%, p NS vs baseline) 30 minutes after the dose. In contrast to single-dose diltiazem, the diltiazem-ibopamine subgroup exhibited an increased left ventricular filling pressure (122%, p < 0.05 vs baseline) and mean pulmonary artery pressure (43%, p < 0.05 vs baseline) at 30 minutes after the dose. One patient experienced a transient episode of chest pain associated with increased heart rate and blood pressure with diltiazem-ibopamine. CONCLUSION: Diltiazem and ibopamine should be coadministered with caution in patients with coronary artery disease and left ventricular dysfunction.

Early ontogeny of immune cells and their functions in the fetal pig.

The origin of immune cells and their products have been studied in the prenatal period in miniature pigs. Macrophages were first detected on day 25, and myelocytes and lymphoid cells by day 28. Membrane antigens SLA-DR and CD45 were found by day 22, membrane molecules MG-7, 8/1, CD1, CD2 and 74-22 by day 28, Gamma/delta T cells were found initially in extrathymic sites (in the liver). The first gamma/delta T cells were detected as early as 40 days of gestation. The expression of fibronectin, Thy-1 and its message, Ig isotypes and the first induction of IFN alpha were described.

Analysis of monoclonal antibodies reactive with the porcine CD2 antigen.

As result of the First International Swine CD Workshop, six monoclonal antibodies (mAbs) (numbers 014, 023, 024, 057, 128, and 130) clustered closely to the internal standard anti-porcine CD2 mAb, MSA4. Despite the close clustering, the cluster was split into two subgroups. To further characterize the relationship between these mAbs, they were used in flow cytometry to inhibit binding of MSA4 to porcine lymphocytes. mAbs 014 (1038-8-31), 023 (MAC83), 024 (MAC80), 057 (PG168), and 128 (MSA4) completely inhibited the binding of MSA4, mAb 130 (MSA2) failed to inhibit MSA4 binding. On dual parameter flow cytometry comparing MSA2 with MSA4, all MSA2+ cells were MSA4+, two thirds of the MSA4+ cells were MSA2-. We conclude that five of the mAbs bind to the same or a closely related epitope on porcine lymphocytes. mAb 130 appears to have aberrantly clustered with the CD2 group of mAb.

Analysis of monoclonal antibodies reactive with the porcine CD4 antigen.

As result of the First International Swine CD Workshop, four monoclonal antibodies (mAb) (#002, 054, 118, and 127) clustered closely to the internal standard anti-porcine CD4 mAb, 74-12-4. To further characterize the relationship between these mAb, they were used in flow cytometry to inhibit binding of 74-12-4 to porcine lymphocytes. mAb #002 (74-12-4) and #054 (PT90) completely inhibited, while mAb #127 (10.2H2) and #118 (b38c6) partially inhibited (57% and 77% respectively), the binding of 74-12-4. Furthermore, none of the mAbs bound to a 74-12-4 negative strain of pigs. We conclude that the four mAb bind to the same, or a closely related, epitope on porcine lymphocytes.

Analyses of monoclonal antibodies reactive with porcine CD5.

Among all monoclonal antibodies (mAbs) analyzed in the first porcine CD workshop, six mAbs showed reactivity to the porcine CD5 antigen (workshop mAbs 067, 068, 069, 070, 071 and 119). Because of lack of immunoprecipitation studies for five (067, 068, 069, 070 and 071) out of the six mAbs, only one mAb (119) could be definitely characterized as mAb against the monomeric 63 kDa porcine CD5 antigen. The five other mAbs included in this cluster are characterized by an identical labelling pattern in FCM and competition of the CD5 epitope recognized by mAb 119. These mAbs were allocated to the wCD5 subcluster.

Analyses of monoclonal antibodies reactive with porcine CD6.

Amongst the monoclonal antibodies (mAbs) submitted to the first porcine CD workshop, two mAbs (workshop numbers 055 and 120) could be identified to recognize the porcine CD6 analogue. Both mAbs seemed to be highly T-cell specific and showed neither reactivity with cells of the myeloic lineage nor with B lymphocytes. The observed molecular mass of the antigen precipitated by mAb 120 of 110 kDa confirmed this classification. Without molecular analyses of the antigen recognized by mAb 055, but similar staining pattern in FCM compared with 120, mAb 055 was allocated to the wCD6 subcluster.

Analyses of mAb reactive with porcine CD8.

Among all mAb submitted to the first porcine CD workshop, based on FCM analyses six mAb could be identified to recognize the porcine CD8 analogue (workshop Nos. 004, 051, 052, 053, 108 and 109). In immunoprecipitation studies three mAb (Nos. 004, 108 and 109) recognized an antigen with an apparent molecular mass of about 35 kDa under reducing conditions and about 70 kDa under non-reducing conditions. The molecular masses of the antigens recognized by the three other mAb (Nos. 051, 052 and 053) are still unknown. Epitope analyses performed by blocking experiments led to the determination of two CD8 epitopes: CD8a and CD8b. CD8a is recognized by mAb Nos. 004, 051 and 052, and CD8b by Nos. 053, 108 and 109.

Analysis of mAb reactive with the porcine SWC1.

Among 54 mAb determined to be reactive with porcine T lymphocytes and/or activation antigens, eight mAb (workshop Nos. 005, 031, 080, 091, 092, 093, 094 and 110) derived from different laboratories grouped together in the T11 cluster and were ordered into the SWC1. One mAb (No. 111) which belong also to this group was lost during the workshop. The SWC1 antigen is a molecule expressed on the majority of leukocytes, resting T lymphocytes, monocytes and granulocytes, but not on B lymphocytes. On T lymphocytes it is down-regulated after activation. The molecular mass of the antigen is unknown. Epitope analyses revealed that seven out of the nine mAb recognized similar epitopes on the SWC1 molecule.

Summary of workshop findings for porcine T-lymphocyte antigens.

Fifty-four mAb preselected in the first round of the first porcine CD workshop for their possible reactivity with T-lymphocyte specific antigens and/or activation antigens were further analysed in a second round. PBMC, thymocytes and nylon-wool purified T lymphocytes derived from peripheral blood, mesenteric lymph nodes and spleen served as target cells for flow cytometric analyses. For the classification of activation antigens several experiments were performed with activated, mitogen-stimulated T lymphocytes and long-term T-lymphocyte cultures. Out of the 54 mAb, 35 mAb could be distributed to six different CD clusters and two swine workshop clusters (SWC). Five mAb could be distributed to the porcine CD2, four mAb to the CD4. Six mAb seemed to recognize the porcine CD5 and two mAb the porcine CD6 analogue. Six mAb were directed against the porcine CD8, whereas two different epitopes could be defined. One mAb was directed against the porcine CD25 analogue. Nine mAb could be clustered to the SWC1, defining an antigen on T lymphocytes and cells of the myeloic linage. Two mAb with high T-cell specificity were clustered to the SWC2.