The combination of X-ray-mediated radiosurgery and gene-mediated immunoprophylaxis for advanced intracerebral gliosarcomas in rats

Rats with advanced, imminently lethal, 4mm diameter, left-sided intracerebral 9L gliosarcoma (9LGS), a well characterized malignant tumor with some similarities to human high-grade astrocytomas, were used as a therapy model 14 days post-implantation of 10⁴ cells. Such tumor-bearing rats die within two weeks (median, 6 days) thereafter if untreated. However, if these tumors are exposed on day 14 to 12–25Gy of an electron-equilibrated 6MV photon beam (radiosurgery), survival is extended about 5–6 fold to a median of 34 days, but long-term survival (> 1 year) is increased only to 18%. Multiple subcutaneous inoculations of radiation-disabled 9LGS cells post-radiosurgery (immunoprophylaxis) extended lifespan and long-term (> 1 year) survival minimally (median, 37 days; 25%, respectively). In sharp contrast, radiosurgery followed by multiple subcutaneous inoculations of radiation-disabled 9LGS cells that had been transfected with granulocyte macrophage colony stimulating factor (GMCSF), a cytokine with demonstrated immune-enhancing properties (i.e. gene-mediated immunoprophylaxis, GMIMPR) increased long-term survival to 67%. To our knowledge, these results are the first to show that the combination of photon radiosurgery and GMIMPR is effective for an advanced, imminently lethal brain tumor in a mammal. These data raise the possibility that GMIMPR following radiation therapy might prove effective for the treatment of some human malignant gliomas.     

Selected adnexal cystic masses in postmenopausal women can be safely managed by laparoscopy

The aim of this study was to assess the efficacy and safety of laparoscopic treatment for adnexal cystic masses that were predicted to be benign in postmenopausal women. Postmenopausal women found to have an adnexal cystic mass were retrospectively evaluated with transvaginal ultrasonography, and serum CA-125 levels. The selection criteria were adnexal cystic masses greater than 3 cm but less than 10 cm, the masses were in the benign range (4-8) of Sassone's scoring system for transvaginal ultrasonography, and the patients had serum CA-125 levels less than 65 IU/mL. Two hundred nineteen women fulfilled the criteria and underwent operative laparoscopy. Almost all the masses (99.5%) were accurately predicted to be benign except for one borderline ovarian tumor. Two hundreds thirteen (97.3%) women were successfully managed by operative laparoscopy and six (2.7%) required laparotomy. For the patients managed by laparoscopy, the mean operative time was 51.3 min; the mean hospital stay was 2.5 days. There was no significant morbidity and surgery-related mortality. The combination of the Sassone's scoring system for transvaginal ultrasonography and serum CA-125 level can accurately predict benign cystic masses, and operative laparoscopy is technically feasible and safe for the management of adnexal mass in postmenopausal women.     

Endoscopic instrumented posterolateral lumbar fusion with Healos and recombinant human growth/differentiation factor-5

OBJECTIVE: The goal of this study was to use a minimally invasive endoscopic surgical technique in a sheep model to evaluate the efficacy of an osteoinductive growth factor, recombinant human growth/differentiation factor-5 (also designated MP52), and an osteoconductive matrix formulation (Healos; DePuy AcroMed, Inc., Mountain View, CA) for inducing and facilitating bone formation. METHODS: Twelve mature sheep underwent bilateral posterolateral lumbar fusion and pedicle screw fixation via a posterior endoscopic approach. Each sheep received two different types of graft material, autogenous iliac crest bone, or a bone graft substitute (MP52 with Healos), inserted into the right and left sides of the spine in an alternating fashion. Groups of four sheep were killed at 2, 4, and 6 months postoperatively for manual, radiographic, and histological evaluation. RESULTS: No neurological impairments, infections, or other complications were observed. After 2 months, partial fusion on both sides was observed, but radiographic evaluation showed greater bone growth on the side that received the bone graft substitute. Solid posterolateral fusion was observed in both autograft and bone graft substitute sites at 4 and 6 months, and autograft and Healos MP52 fusion sites were essentially the same at histological examination. There was no abnormal overgrowth of new bone from either of these two materials. CONCLUSION: Endoscopic posterolateral lumbar arthrodesis and instrumentation is feasible, safe, and effective in a sheep model. Healos is a useful bonding agent that mimics natural bone in posterolateral intertransverse fusion sites. Combined with MP52, it produced fusion comparable to that of autogenous bone graft. Minimally invasive techniques and bone graft substitutes could eliminate morbidity and increase the likelihood of successful fusion.     

RU486 blocks fasting-induced decrease of neuronal nitric oxide synthase in the rat paraventricular nucleus

It has been reported that food deprivation decreases expression of neuronal nitric oxide synthase (nNOS) in the hypothalamic paraventricular nucleus (PVN). Food deprivation produces autonomic changes and the PVN nitric oxide has been suggested to be involved in regulation of autonomic functions. In order to understand the molecular mechanism by which food deprivation decreases nNOS expression in the PVN, we examined if plasma glucocorticoids, which reported to be elevated during food deprivation, mediates the fasting-induced down-regulation of the PVN-nNOS. Male Sprague-Dawley rats underwent 48 h of food deprivation, but not water deprivation, with/without subcutaneous RU486, glucocorticoid receptor antagonist, and the brain tissues were processed for immunohistochemistry with specific antibodies against nNOS. Immunoreactivity of phosphorylated cAMP response element-binding protein (pCREB) was also examined in the PVN sections, because nNOS promoter carries cAMP response element (CRE). Food deprivation significantly decreased both nNOS and pCREB immunoreactivity (-ir) in the medial parvocellular PVN, and RU486 blocked this reduction. In the posterior magnocellular PVN, nNOS-ir, but not pCREB-ir, was decreased by food deprivation, and RU486 exerted no effect. These results suggest that glucocorticoid receptor may mediate the fasting-induced down-regulation of nNOS in the parvocellular PVN, but not in the magnocellular PVN.     

Induction of cytochrome P450s by rutaecarpine and metabolism of rutaecarpine by cytochrome P450s

Rutaecarpine is an alkaloid originally isolated from the unripe fruit of Evodia rutaecarpa. Recently, rutaecarpine has been characterized to have an anti-inflammatory activity through cyclooxygenase-2 inhibition. In the present studies, the effects of rutaecarpine on liver cytochrome P450 s (P450s) and P450 s involved in the metabolism of rutaecarpine were studied in vivo and in vitro, respectively, because the data are crucial in the early development of rutaecarpine as a new drug candidate. Oral administration to male ICR mice of rutaecarpine for 3 consecutive days induced liver P450 1A-, 2B- and 2E1-selective monooxygenase activities. The induction of P450 1A and 2B by rutaecarpine was confirmed by Western immunoblotting. When rutaecarpine was incubated with rat liver microsomes in the presence of an NADPH-generating system, five metabolites were detected by UV and mass spectral analyses. The 3-methylcholanthrene- and phenobarbital-induced microsomes greatly increased the formation of metabolites. Our present results suggest that rutaecarpine might induce P450 1A and 2B in mice, and that P450 1A and 2B might predominantly metabolize rutaecarpine in rat liver microsomes.     

Selective effects of light exposure on distribution of motility in the chick embryo at E18

It is well established that orderly patterns of motor neuron activity, muscle recruitment, and limb movement are generated in chicks during motility by embryonic day (E)9, the midpoint in embryonic development. However, our recent work suggests that some attributes of motility, such as the rhythm of repetitive limb movements and distribution of activity, become less orderly after E9. In this study, we extend these observations by performing continuous force recordings over a 24-h period in ovo at E18 with augmented sampling of synchronized video and electromyogram (EMG) recordings. We report the distribution of three repetitive behaviors, rapid limb movement, respiratory-like movement, and beak clapping, identified in force recordings, and the general distribution of motility. We also test a model recently proposed to account for age-related changes in motility parameters. In the model, we proposed that circadian networks contribute to the age-related changes in distribution of motility. As a first test of this hypothesis, we examine whether light exposure contributes to the variable distribution of motility by comparing motility parameters at E18 for embryos incubated and tested under either a 12-h light/dark cycle or continuous light. Results suggest that exposure to light increases the total amount of activity and hastens the onset of extended respiratory-like movement sequences but does not impact expression of repetitive limb movement or beak clapping at E18. The possible influence of circadian mechanisms on embryonic behavior and insensitivity of repetitive limb movements to light exposure are discussed.     

Acute chorioamnionitis and intra-amniotic inflammation are more severe according to outside-in neutrophil migration within the same chorio-decidua

ObjectiveNo information exists about whether acute histologic chorioamnionitis (acute-HCA) is more advanced and severe, and intra-amniotic inflammation is more frequent and intense according to outside in neutrophil migration within the same chorio-decidua. The objective of current study is to examine this issue.Materials and methodsWe included 106 singleton preterm-births (gestational age at delivery: 20–34 weeks) due to either preterm-labor or preterm-PROM in the context of acute chorio-deciduitis. Study-population was divided into 3 groups according to outside-in neutrophil migration within chorio-decidua as follows: 1) group-1: ‘inflammation restricted to the decidua’ (n = 22); 2) group-2: ‘inflammation restricted to the MT of chorion and the decidua’ (n = 31); 3) group-3: ‘inflammation in the CT of chorion’ (n = 53). We examined the frequency of inflammation in each placental compartment beyond chorio-decidua (i.e., amnion, umbilical cord, and chorionic-plate), and total grade (1–8) of acute-HCA. Moreover, the frequency of intra-amniotic infection (defined as positive amniotic-fluid culture for aerobic and anaerobic bacteria and genital mycoplasmas) and intra-amniotic inflammation (defined as amniotic fluid WBC ≥ 19 cells/mm³), and an intra-amniotic inflammatory response gauged by amnioticfluid WBC count (cells/mm³) were examined in 50 amniotic fluid samples within 7 days of birth.ResultsAmnionitis, funisitis and chorionic plate inflammation were more frequent (each for P < 0.01) and median total grade of acute-HCA was increased (P < 0.001) according to outside-in neutrophil migration within chorio-decidua (group-1vs.group-2vs.group-3). Moreover, intra-amniotic infection and inflammation were more frequent (each-for P < 0.05) and median amniotic-fluid WBC count was increased (P < 0.01) according-to outside-in neutrophil-migration within chorio-decidua (group-1 vs. group-2 vs. group-3).ConclusionAcute-HCA is more advanced and severe, and intra-amniotic inflammation is more frequent and intense according to outside in neutrophil migration within the same chorio-decidua. This finding suggests that what is now acute chorio-deciduitis should be subdivided.     

Synthesis and biological activity of conformationally controlled 2-PAM derivatives

A series of conformationally controlled 2-PAM derivatives were prepared from 2-acetylpyridine and 2,3-pyrido[b]cycloalkenones in two steps and their reactivities towards parathion poisoned AChE were evaluated. The most planar 2,3-pyrido[b]cyclohexanone oxime methiodide showed an activity comparable to 2-PAM implying E-syn is that the most active conformation of 2-PAM in the biological system.