Inhibition of neutrophil and monocyte recruitment by endogenous and exogenous lipocortin 1

1. The role played by endogenous lipocortin 1 in the anti-migratory action exerted by dexamethasone (Dex) on monocyte recruitment in an in vivo model of acute inflammation was investigated by use of several neutralizing polyclonal antibodies raised against lipocortin 1 or a lipocortin 1-derived N-terminus peptide (peptide Ac2-26). The efficacy of peptide Ac2-26 in inhibiting monocyte and polymorphonuclear leucocyte (PMN) recruitment was also tested.
2. Intraperitoneal (i.p.) injection of zymosan A (1 mg) produced a time-dependent cell accumulation into mouse peritoneal cavities which followed a typical profile of acute inflammation: PMN influx was maximal at 4 h post-zymosan (between 15 and 20×10⁶ cells per mouse), and this was followed by an accumulation of monocytes which peaked at the 24 h time-point (between 10 and 15×10⁶ cells per mouse).
3. Dex administration to mice reduced zymosan-induced 4 h PMN infiltration and 24 h monocyte accumulation with similar efficacy: approximately 50% of inhibition of recruitment of both cell types was achieved at the dose of 30 μg per mouse (∼1 mg kg⁻¹, subcutaneously (s.c.)). Maximal inhibitions of 64% and 67% on PMN and monocyte recruitment, respectively, were measured after a dose of 100 μg per mouse (∼3 mg kg⁻¹, s.c.).
4. Dex (30 μg s.c.) inhibited monocyte (53%) and PMN (69%) accumulation in response to zymosan application in mice which had been treated with a non-immune sheep serum (50 μl s.c.). In contrast, the steroid was no longer active in reducing cell accumulation in mice which had been passively immunized against full length human recombinant lipocortin 1 (serum LCS3), or against lipocortin 1 N-terminus peptide.
5. Treatment of mice with vinblastine (1 mg kg⁻¹, intravenously (i.v.)) produced a remarkable leucopenia as assessed 24 h after administration. This was accompanied by a 60% reduction in 4 h-PMN influx, and by a 27% reduction in 24 h-monocyte accumulation, measured after zymosan administration. The inhibitory effect of Dex on monocyte recruitment was not significantly modified in vinblastine-treated mice, with 36% and 57% of inhibition calculated at the dose of 30 μg Dex, and 70% and 60% of inhibition at 100 μg Dex, in vehicle- and vinblastine-treated mice, respectively.
6. Treatment of mice with peptide Ac2-26 dose-dependently attenuated PMN influx at 4 h post-zymosan with a significant effect at 100 μg per mouse (45% of inhibition, n=9, P<0.05) and a maximal effect of 61% inhibition at the highest dose tested of 200 μg s.c. (n=14, P<0.05). No effect of peptide Ac2-26 (200 μg s.c.) was seen on zymosan-induced 24 h monocyte recruitment. In contrast, administration of 200 μg peptide Ac2-26 every 6 h was effective in reducing the number of monocytes harvested from the inflamed peritoneal cavities at 24 h post-zymosan: 9.40±0.58×10⁶ monocytes per mouse (n=13) and 5.74±0.34 monocytes per mouse (n=14) in vehicle- and peptide Ac2-26-treated mice, respectively (P<0.05).
7. Finally, peptide Ac2-26 produced a concentration-dependent inhibition of the rate of phagocytosis of mouse resident peritoneal macrophages as measured by flow cytometry, with a maximal reduction of 34% at the highest concentration tested of 100 μg ml⁻¹ (n=8 experiments performed in duplicate; P<0.05).
8. In conclusion, this study suggests that in vivo monocyte recruitment during acute inflammation is, at least in part, under the negative modulatory control of endogenous lipocortin 1 (as seen after administration of Dex by using the specific antisera) and exogenous lipocortin 1 mimetics (as observed with peptide Ac2-26). In addition to the neutrophil, we can now propose that the monocyte also can be a target for the in vivo anti-inflammatory action of lipocortin 1.

     

Xanthogranulomatous pyelonephritis

A four years-old boy with Xanthogranulomatous pyelonephritis was surgically treated at the Pediatric Surgery Unit of the Santa Casa de Misericórdia of Maceió. Comments are made upon pathology, pre-operative diagnostic difficulties, differential diagnosis, and the rare occurrence in children.     

Prognostic factors in fetal hydronephrosis: a multivariate analysis

With the increasing use of obstetric echography fetal hydronephrosis has been reported more frequently. The purpose of this study was to identify prognostic factors associated with adverse outcome, such as renal failure and death, in fetal hydronephrosis. One hundred and forty-eight children with fetal hydronephrosis were admitted, submitted to a systematic protocol, and prospectively followed. Prognostic factors associated with fetal echography and clinical and laboratory findings on admission were studied. The median follow-up was 39 months. The analysis was conducted in two steps. In a univariate analysis, variables associated with adverse outcome were identified by the Kaplan-Meier method. The variables that were significantly associated with adverse outcome were then included in a multivariate analysis. This analysis, using the multivariate Cox’s model, was performed to identify variables that were independently associated with a worse prognosis. Only variables that remained independently associated with adverse outcome were included in the final model. After final adjustment by Cox’s multivariate model, three variables were identified as independent predictors of adverse outcome: oligohydramnios, prematurity, and glomerular filtration rate lower than 20 ml/min. Thus, in the presence of oligohydramnios, prematurity, and abnormal renal function, the medical team must plan appropriate follow-up for infants at health centers prepared to investigate and treat uropathies in newborns. Received: 24 August 1998 / Revised: 7 December 1998 / Accepted: 11 December 1998     

Prevention by rolipram of concanavalin A-induced T-cell-dependent hepatitis in mice

Rolipram is a type IV phosphodiesterase inhibitor endowed with powerful immunomodulatory properties. In this study, we evaluated the effects of this drug on the development of the T-cell-mediated hepatitis inducible in mice by concanavalin A. The results indicated that prophylactic treatment with either 5 or 10 mg/kg rolipram injected intraperitoneally 24 h and 1 h prior to intravenous (i.v.) challenge with 20 mg/kg concanavalin A successfully ameliorated serological and histological signs of liver damage, so that the treated mice showed lower transaminase levels in the plasma and milder mononuclear cell infiltration of the liver as compared to vehicle-treated controls. Moreover, this effect was associated with profound modifications of circulating levels of cytokines released after concanavalin A injection, with the blood levels of interferon-gamma and tumor necrosis factor-alpha being significantly lower and those of interleukin-10 higher than those of the control mice. In particular, the increased blood levels of interleukin-10 might play an important role in the anti-hepatitic effects of rolipram as coadministering this compound with anti-interleukin-10 monoclonal antibody significantly reduced its anti-inflammatory action. These results suggest that rolipram may be useful in the clinical setting for the treatment of cell-mediated immunoinflammatory diseases such as immunoinflammatory hepatitis.     

Experimental brain tumors by transplacental ENU

Summary Experimental cerebral tumors have been induced by transplacental ENU. The morphologic study of the brains of treated rats revealed that cellular hyperplasias appear at the 30th day of extrauterine life in the paraventricular white matter, i.e., before the already known early neoplastic proliferations. Cytofluorimetric investigations failed to demonstrate differences between treated and control rats during the 1st month. On the contrary, adenylate cyclase activity is very high in that period. The duration of the latency period is discussed.This research was supported by Grants No. 79.00678.96 and No. 79.00664.96 of Progetto Finalizzato Control of Neoplastic Growth, Consiglio Nazionale delle Ricerche (C.N.R.), Rome     

Invivo control of tumor-growth by lonidamine and radiations - influence of the timing and sequence of drug administration and irradiation treatment

It has been well established that Lonidamine (LND), [1,(2,4 dichlorobenzyl)-1H-indazol-3-carboxylic acid], affects tumor growth and enhances the effect of X-ray both in vitro and in vivo. Nevertheless, difficulties arise if the available experimental data should be utilized to design clinical trials since schedules, routes of administration as well as dosages greatly differ from those currently employed in the clinic. With the aim to overcome these difficulties, experiments with modalities similar to those employed in the current clinical practice have been undertaken to evaluate: (i) the influence of the LND dosage on the antitumor effect; (ii) the time lenght of its administration for the optimal effect; (iii) the best schedule of treatment when LND is associated with radiations. The results may be summarized as follows: (i) antitumor effectiveness of LND, in terms of growth delay, increases with LND dosage. Moreover, the drug administered from the day of transplant significantly decreases the tumor takes. (ii) to exert the antineoplastic effect LND must be administered continuosly because if the treatment is interrupted the tumor regrows like an untreated one. (iii) the maximal response of the association X-ray-LND is elicited when the drug is given after irradiation treatment.     

Radiological Investigations in Vertical Banded Gastroplasty Patients

In our Institute we have performed 124 vertical banded gastroplasties. Patients with a follow-up beyond 3 months were studied with a barium meal, in order to evaluate the efficiency of surgery and the eventual complications. Seventy-nine patients have had one or more X-ray investigations at various times after surgery (for a total of 136 studies). The first 20 patients were routinely studied at 1, 2 and 3 years after the operation; the next 32 patients were studied for features such as vomiting, poor weight loss or low food intake; the last 27 patients were studied with an early overlook beginning 3 months after surgery. We noted gastroesophageal reflux in eight (10.1%) cases, outlet dilatation in four (5%) cases, outlet substenosis (diameter 6-8 mm) in 13 (16.4%) cases, outlet stenosis (diameter ≤5 mm) in four (5%) cases, peanut-type deformation in three (3.7%) cases, and staple-line disruption in 17 (21.5%) cases. The staple-line disruption was correlated in the first part of the series with a reinforcement of such a suture, while the last 27 patients, with vertical stapling carried out with a 4-row stapler without reinforcement, did not present any disruption. The radiographic examination gives information about weight loss and side effects.     

喉鳞状细胞癌组织体外原代培养的初步研究

目的 应用体外培养技术,对喉鳞状细胞癌组织进行体外培养,探讨喉鳞癌组织原代培养中的各种影响因素,为建立人喉鳞癌组织的细胞系提供实验基础。方法采用体外组织培养技术,对24例人喉鳞状细胞癌组织进行原代培养,观察原代培养中肿瘤细胞的生长与供体的年龄、肿瘤组织的分化程度及不同培养方法的关系,分析在人喉鳞癌细胞的培养中成纤维细胞、微生物污染的影响。结果 24例人喉鳞状细胞癌组织标本,年龄小于60岁组的细胞生长率为31.25％(5/16例),年龄大于60岁组为37.5％(3/8例);高分化组为100％(2/2例),中分化组为30.8％(4/13例),低分化组25％(2/8例);组织块培养法为43.75％(7/16例),酶消化法培养为10％(1/10例);倒置显微镜下观察,在培养的第5～7天,在贴壁组织块周围可见到有上皮样细胞爬出。全部标本中,成纤维细胞的过度生长和微生物的污染是阻碍人喉鳞癌细胞生长的重要因素。结论培养组织的细胞生长率与供体的年龄关系不大;肿瘤组织的分化程度较高者,细胞的生长率较高;与酶消化分离培养法相比,贴壁组织块培养法的细胞生长率较高;成纤维细胞及微生物的污染是阻碍人喉鳞癌细胞系建立的重要因素。     

Propofol and remifentanil in day surgery

BACKGROUND: Aim of the study: to compare AG versus MAC using propofol & remifentanil in a day surgery setting evaluating intra and postoperative clinical conditions and emergence times. METHODS: Propofol and remifentanil, either for general anesthesia (AG) then conscious sedation (MAC), have been administered to 218 patients undergoing mainly plastic or proctologic surgery as day hospital. AG was induced with propofol 1.5-2 mg/kg followed by a continuous infusion of 10 mg/kg/h and remifentanil infused at 10 micrograms/kg/h; MAC was started with propofol 3 mg/kg/h and remifentanil 4-5 micrograms/kg/h; during the maintenance phase of both AG and MAC, infusion rates of both drugs were adjusted according to clinical needs. Diazepam (0.05-0.06 mg/kg) and/or midazolam (2-3 mg) were given as premedication or coinduction as necessary. All patients received field infiltration with local anesthetics (lidocaine or mepivacaine); patients under GA were artificially ventilated with O2/air through IOT or LMA. Surgical and anesthesiological data were collected on specially designed records, with special attention to time intervals between anesthesia (FA) and surgery (FC) end and eyes opening (EO), orientation (OR), return of spontaneous breathing (SR), extubation (EST), sitting (SED), walking (CAMM), dressing (VEST) and discharge (DIM); data were analyzed with parametric and non parametric analysis of variance. RESULTS: All emergence intervals were longer under AG than under MAC: the earlier in the range of 4-5 vs 0.5-1 min; for the late intervals; FA-SED 24 +/- 18 vs 15 +/- 8, FA-PIED 65 +/- 48 vs 34 +/- 17, FA-VEST 69 +/- 58 vs 33 +/- 17, FA-CAMM 68 +/- 42 vs 39 +/- 19. Discharge times (83 +/- 67 vs 73 +/- 60) were similar between the two groups. Drugs consumption under AG were roughly double than under MAC; total dose infused of propofol (mg/kg/min) 0.118 +/- 0.044 vs 0.06 +/- 0.036; total dose of remifentanil (microgram/kg/min): 0.106 +/- 0.049 vs 0.066 +/- 0.027. AG resulted in a higher % incidence of intraoperative hypotension and bradycardia: hypotension 61.7 vs 25.7 and bradycardia 30.3 vs 12.4. SaO2 decreased more commonly during MAC than AG (20.9% vs 10.1); intraoperative itching was referred in 20% of MAC patients. Conversions rate from MAC to AG was 2.8%. Psychomotor agitation was more frequent following AG (14%) than MAC (2%); nausea (1%), vomiting, shivering (12%), headache (2%), ortostatic hypotension (2%) were similar between the two groups. Diazepam and/or midazolam caused a significant prolongation of recovery intervals, for both AG and MAC with a mean delay of the order of 100-200%. CONCLUSIONS: Propofol- remifentanil gave excellent conditions for a wide variety of day surgery procedures, offering good anesthesia with quick emergence; the addition of bdz, even at low doses, prolongs significantly discharge times.