Craniospinal irradiation with concurrent temozolomide for primary metastatic pediatric high-grade or diffuse intrinsic pontine gliomas. A first report from the GPOH-HIT-HGG Study Group.

BACKGROUND: High-grade (HGG) and diffuse intrinsic pontine gliomas (DIPG) with primary metastatic spread are extremely rare and have a dismal prognosis. Analogous to simultaneous radiochemotherapy in non-metastatic HGG and DIPG, concurrent craniospinal irradiation (CSI) and metronomic temozolomide (metroTMZ) may represent a reasonable therapeutic approach. However, the antitumor efficacy and toxicity of this treatment still have to be investigated. PATIENTS AND METHODS: Between March 2007 and December 2012, six children with primary metastatic HGG (n = 4) or DIPG (n = 2) received CSI and concurrent metroTMZ based on individual treatment recommendations and, in some cases, within the HIT-HGG 2007 multicenter trial. Outcome and treatment-related toxicities were evaluated. RESULTS: All patients received irradiation to the entire craniospinal axis (35.2 Gy, n = 5; 36 Gy, n = 1:) and 5 received a local boost to macroscopic tumor deposits. Simultaneously, metroTMZ (75 mg/m(2)/day, n = 5; 60 mg/m(2)/day, n = 1) was administered. Additionally, 1 patient received nimotuzumab once per week. Within a median follow-up of 10.0 months (range 6.5-18.7 months), all patients experienced disease progression and 5 patients died. Median progression-free survival was 4.0 ± 0.8 months (range 2.4-10.7 months) and median overall survival was 7.6 ± 3.5 months (range 4.0-17.6 months). Acute myelosuppression most severely limited application of this aggressive treatment strategy. Severe hematotoxicities (≥ grade 3) occurred in all patients and metroTMZ had to be interrupted or discontinued in 4 out of 6 cases. CONCLUSION: Concurrent CSI and metroTMZ might represent a feasible treatment approach for primary metastatic HGG and DIPG. On the basis of our experience, severe but manageable acute hematotoxicity has to be expected. An international effort is warranted to reassess the efficacy and toxicity of this approach within a prospective study.

Expression of annexin II in conventional renal cell carcinoma is correlated with Fuhrman grade and clinical outcome.

Conventional renal cell carcinomas (CRCCs) were investigated for the expression of annexin II (ANX II) to determine out whether this calcium-binding protein could serve as a useful prognostic marker. CRCCs and adjacent nonneoplastic tissue from 33 patients were investigated for ANX II by immunohistochemistry, RT-PCR, and western blot analysis. ANX II expression was correlated with tumor differentiation (Fuhrman grade) and to clinical outcome. Tumors were composed of ANX II positive and negative cells. In grade I tumors only a weak membranous staining was seen in immunopositive cells. In grade II and III tumors, however, ANX II was seen in the cytoplasm and at the cell membranes of tumor cells. On serial sections membranous and cytoplasmic immunoreactivity for ANX II occurred predominantly in eosinophilic cells whereas clear cells were mostly immunonegative. The ANX II expression in CRCCs was correlated with clinical outcome and Fuhrman grade. Since ANX II expression is correlated with Fuhrman grade and clinical outcome it may be a useful marker for prognosis in CRCC.

Two hydrophobic protein fractions of ovine pulmonary surfactant: isolation, characterization, and biophysical activity.

Pulmonary surfactant contains two extremely hydrophobic proteins, SP-B and SP-C. We present a novel HPLC method for the preparation of these hydrophobic proteins. It is based on size-exclusion chromatography using the apolar stationary-phase butyl silica gel and isocratic elution with acidified chloroform/methanol. Samples for HPLC were prepared from sheep lung lavage fluid by centrifugation and extraction with chloroform/methanol. Amino acid analyses of the two protein fractions revealed sequences that are consistent with SP-B and SP-C, respectively. MALDI-TOF-MS analyses of the SP-B fraction showed one major peak of dimeric SP-B with m/z 17,361, and additional peaks of monomeric and oligomeric forms, which are predominantly even numbered. The SP-C fraction showed a peak at m/z 4200, consistent with the theoretical mass of the dipalmitoylated form of this protein. The biophysical activity of pure sheep SP-B and SP-C was evaluated by measuring the surface tension using axisymmetric drop shape analysis for captive bubbles. We found distinct surface pressure versus surface area isotherms of SP-B and SP-C indicating different biophysical activities for these surfactant proteins. The new preparative HPLC method is able to replace the established, time-consuming low-pressure liquid chromatography method for the isolation of SP-B and SP-C from lipids.

Folding incompetence of cathepsin L-like cysteine proteases may be compensated by the highly conserved, domain-building N-terminal extension of the proregion.

Folding of cathepsins L and S depend upon their proregion which extends the enzyme part by about 100 amino acids. Only a minority of the prosequence follows the structural template provided by the enzyme part; the majority forms an autonomous minidomain fairly distant from the active site cleft. We suggest that this prodomain may be the structural correlate of a foldase function of the proregion within the cathepsin L-like subfamily of papain-type cysteine proteases and report on a functional approach supporting this hypothesis.

Polymorphisms of human SP-A, SP-B, and SP-D genes: association of SP-B Thr131Ile with ARDS.

An allele association study of 19 polymorphisms in surfactant proteins SP-A1, SP-A2, SP-B, and SP-D genes in acute respiratory distress syndrome (ARDS) was carried out. Trend-test analysis revealed differences (p < 0.05) in the frequency of alleles for some of the microsatellite markers flanking SP-B, and for one polymorphism (C/T) at nucleotide 1580 [C/T (1580)], within codon 131 (Thr131Ile) of the SP-B gene. The latter determines the presence or absence of a potential N-linked glycosylation site. Multivariate analysis revealed significant differences only for the C/T (1580) polymorphism. When the ARDS population was divided into subgroups, idiopathic (i.e., pneumonia, etc.) or exogenic (i.e., trauma, etc.), significant differences were observed for the C/T (1580), for the idiopathic ARDS group, and the frequency of the C/C genotype was increased in this group. Based on the odds ratio, the C allele may be viewed as a susceptibility factor for ARDS. Although the expression of both C and T alleles occurs in heterozygous individuals, it is currently not known whether these alleles correspond to similar levels of SP-B protein. These data suggest that SP-B or a linked gene contributes to susceptibility to ARDS.

Unilateral lung edema: effects on pulmonary gas exchange, hemodynamics, and pulmonary perfusion distribution.

Two types of unilateral lung edema in sheep were characterized regarding their effects on pulmonary gas exchange, hemodynamics, and distribution of pulmonary perfusion. One edema type was induced with aerosolized HCl (0.15 M, pH 1.0) and the other with NaCl (0.15 M, pH 7.4). Both aerosols were nebulized continuously for 4 h into left lungs. In HCl-treated animals, pulmonary gas exchange deteriorated [from a partial arterial O(2) pressure-to-inspired O(2) fraction ratio (Pa(O(2))/FI(O(2))) of 254 at baseline to 187 after 4 h HCl]. In addition, pulmonary artery pressure and total pulmonary vascular resistance increased (from 16 to 19 mmHg and from 133 to 154 dyn. s. cm(-5), respectively). In NaCl-treated animals, only the central venous pressure significantly increased (from 7 to 9 mmHg). Distribution of pulmonary perfusion (measured with fluorescent microspheres) changed differently in both groups. After HCl application, 6% more blood flow was directed to the treated lung, whereas, after NaCl, 5% more blood flow was directed to the untreated lung. HCl and NaCl treatment both induce an equivalent lung edema, but only HCl treatment is associated with gas exchange alteration and tissue damage. Redistribution of pulmonary perfusion maintains gas exchange during NaCl treatment and decreases it during HCl inhalation.

[Effect of preoperative preoperative visualization therapy on postoperative outcome after inguinal hernia surgery and thyroid resection]. / Einfluss einer präoperativen Visualisierungstherapie auf die postoperativen Ergebnisse nach Leistenherniotomie und Strumaresektion.

UNLABELLED: This randomized controlled trial aims to evaluate the influence of preoperative relaxation techniques on postoperative outcomes. From January 1997 to June 1998 208 patients were operated on for primary inguinal hernia or goiter. The patients were randomized into two groups: Group A (n = 103) underwent the surgical treatment with a preoperative visualization therapy. Group B (n = 105) underwent the surgical treatment without a preoperative therapy. Patients with age under 18 years, ASA-status IV-V, recurrent inguinal hernia or recurrent goiter and malignant neoplasms were excluded from the study. There were no differences in age, sex, duration of the operation, training of the surgeon, and preoperative blood parameters between the two groups. RESULTS: During the postoperative follow-up we observed more hematomas (group A with visualization therapy: 30.3%, group B without visualization therapy: 44.4%) as well as more pain (group A: 4.2, group B: 5.2) and analgesic consumption (group A: 59.7 mg Tramadol HCL, group B: 72.5 mg Tramadol HCL) in group B (p < 0.05). There were no significant differences in infections, nausea, hypocalcemia, tetania, recurrent nerve palsy, fever. CONCLUSIONS: Preoperative visualization therapy reduces significantly the number of postoperative hematomas after inguinal hernia repair. Furthermore a decrease of analgesic requirements after surgical treatment was observed.

cDNA cloning of ovine pulmonary SP-A, SP-B, and SP-C: isolation of two different sequences for SP-B.

Pulmonary surfactant promotes alveolar stability by lowering the surface tension at the air-liquid interface in the peripheral air spaces. The three surfactant proteins SP-A, SP-B, and SP-C contribute to dynamic surface properties involved during respiration. We have cloned and sequenced the complete cDNAs for ovine SP-A and SP-C and two distinct forms of ovine SP-B cDNAs. The nucleotide sequence of ovine SP-A cDNA consists of 1,901 bp and encodes a protein of 248 amino acids. Ovine SP-C cDNA contains 809 bp, predicting a protein of 190 amino acids. Ovine SP-B is encoded by two mRNA species, which differ by a 69-bp in-frame deletion in the region coding for the active airway protein. The larger SP-B cDNA comprises 1,660 bp, encoding a putative protein of 374 amino acids. With the sequences reported, a more complete analysis of surfactant regulation and the determination of their physiological function in vivo will be enabled.

Host defence capacities of pulmonary surfactant: evidence for 'non-surfactant' functions of the surfactant system.

The most well characterized function of pulmonary surfactant is its ability to reduce surface tension at the alveolar air-liquid interface, thereby preventing lung collapse. However, several lines of evidence suggest that surfactant may also have 'non-surfactant' functions: specific components of surfactant (proteins and phospholipids) may interact with different alveolar cells, inhaled particles and micro-organisms modulating pulmonary host defence systems. SP-A, the most abundant surfactant protein, binds to alveolar macrophages via a specific surface receptor with high affinity [128]. Such binding effects the release of reactive oxygen species from resident alveolar macrophages if SP-A is properly presented to the target cell. SP-A also stimulates chemotaxis of alveolar macrophages [142], and serves as an opsonin in the phagocytosis of herpes simplex virus [161] Candida tropicalis [138] and various bacteria [137]. In addition, SP-A enhances the uptake of particles by monocytes and culture-derived macrophages [140] and improves bacterial killing. SP-D, another hydrophobic surfactant-associated protein, might interact with alveolar macrophages as well, stimulating the release of oxygen radicals [148], while for the hydrophilic surfactant proteins SP-B and SP-C no macrophage interactions have been described so far. SP-A and SP-D are members of the so-called 'collectins', pattern recognition molecules involved in first line defence. While some surfactant proteins appear to stimulate certain macrophage defence functions, surfactant phospholipids seem to inhibit those of lymphocytes. Suppressed lymphocyte functions include lymphoproliferation in response to mitogens and alloantigens, B cell immunoglobulin production and natural killer cell cytotoxicity. Concerning surfactant's phospholipid composition phosphatidylglycerol is more suppressive than phosphatidylcholine on a molar basis [38]. Bovine surfactant has an immunosuppressive effect on the development of hypersensitivity pneumonitis in a guinea pig model [150]. Despite these interesting observations, several important questions concerning the interactions of surfactant components with pulmonary host defence systems remain unanswered. Sufficient host defence in the lungs works through various humoral-cellular systems in conjunction with the specific anatomy of the airways and the gas exchange surface--how does the surfactant system fit into this network? Surfactant and alveolar cells are both altered during lung injury--is there a relationship between alveolar cells from RDS patients and the endogenous surfactant isolated from such patients? How does exogenous surfactant as used for substitution therapy modulate the defence system of the host? Some of those artificial surfactants have been shown to inhibit the endotoxin-alveolar macrophages, PMNs and monocytes including IL-1, IL-6 and TNF [139,152].(ABSTRACT TRUNCATED AT 400 WORDS)

Compartment-specific immunolocalization of conserved epitopes of the glycoprotein gB of herpes simplex virus type 1 and bovine herpes virus type 2 in infected cells.

Monoclonal antibodies directed against a surface glycoprotein of the bovine herpes virus type 2 (BHV-2, bovine herpes mammillitis virus) recognize also determinants of the major glycoprotein gB of the human herpes simplex virus type 1 (HSV-1). Cross-reacting antigens of the virions and in infected cells were localized with immunocytochemical methods, immunofluorescence as well as pre-embedding and cryoultramicrotomy immune electron microscopy. All antibodies stain to different degrees cell free BHV-2 and HSV-1 virions. In the cell two predominant staining patterns could be observed indicating that expression of epitopes is dependent upon the cell compartment: (i) staining of cytoplasmic membranes and enveloped particles within membrane systems and (ii) staining of intranuclear antigens. Antibodies tagging intranuclear antigens react with moderately dense material or with the periphery of nucleocapsids. This unexpected result is interpreted in terms of two hypotheses: (1) presence of common epitopes on two entirely different herpesvirus proteins conserved in HSV-1 and BHV-2 and (2) transport of gB or its precursor into the nucleus.

Monoclonal antibodies against herpes simplex virus type 1-infected nuclei defining and localizing the ICP8 protein, 65K DNA-binding protein and polypeptides of the ICP35 family.

The production and properties of monoclonal antibodies raised against herpes simplex virus type 1 (HSV-1)-infected cell nuclei are described. Biological and immunochemical assays revealed that these antibodies recognize four different proteins in HSV-1-infected cells. Four antibodies reacted with the major DNA-binding protein (ICP8) and six with the 65K DNA-binding protein. Two antibodies detected the ICP35 family of proteins and one antibody bound to a protein with an apparent mol. wt. of 60K. Immune electron microscopy showed that the major DNA-binding protein had a patchy distribution, whereas the 65K DNA-binding protein was evenly spread in the infected cell nuclei. The 60K protein as well as the polypeptides of the ICP35 family were preferentially found associated with the viral capsid.