Fluid Protein Research Articles

Flow-injection determination of sulfanilamides in drugs and biological fluids with spectrophotometric detection

Operating conditions were found for the determination of sulfanilamide pharmaceuticals such as sulfathiazole, sulfadimezine, ethazole, sulfamethoxypyridazine, sulfadimethoxine, sodium sulfacyl, sulfalene, sulfaguanidine, and carbutamide as 4,6-dinitrobenzofuroxane derivatives by flow-injection analysis with spectrophotometric detection. The best results were obtained when flows of ethanol (methanol) and a buffer solution were mixed at a volume ratio of 30 : 70 and pH 6.5–7.0. The calibration range for the pharmaceuticals was 0.25–5.5 μg/mL. The limits of detection were 0.12–0.24 μg/mL. Sulfanilamides were determined in medicinal forms (tablets, ointments, and drops) and biological fluids (protein hydrolyzate, blood plasma, and whole blood).

TEMPORAL CHANGES IN RESPIRATORY DYNAMICS IN MICE EXPOSED TO PHOSGENE

One hallmark of phosgene inhalation toxicity is the latent formation of life-threatening, noncardiogenic pulmonary edema. The purpose of this study was to investigate the effect of phosgene inhalation on respiratory dynamics over 12 h. CD-1 male mice, 25-30 g, were exposed to 32 mg/m 3 (8 ppm) phosgene for 20 min (640 mg min/m 3) followed by a 5-min air washout. A similar group of mice was exposed to room air for 25 min. After exposure, conscious mice were placed unrestrained in a whole-body plethysmograph to determine breathing frequency (f), inspiration (Ti) and expiration (Te) times, tidal volume (TV), minute ventilation (MV), end inspiratory pause (EIP), end expiratory (EEP) pause, peak inspiratory flows (PIF), peak expiratory flows (PEF), and a measure of bronchoconstriction (Penh). All parameters were evaluated every 15 min for 12 h. Bronchoalveolar lavage fluid (BALF) protein concentration and lung wet/dry weight ratios (W/D) were also determined at 1, 4, 8, and 12 h. A treatment × time repeated-measures two-way analysis of variance (ANOVA) revealed significant differences between air and phosgene for EEP, EIP, PEF, PIF, TV, and MV, p ≤ .05, across 12 h. Phosgene-exposed mice had a significantly longer mean Ti, p ≤ .05, compared with air-exposed mice over time. Mice exposed to phosgene showed marked increases (approximately double) in Penh across all time points, beginning at 5 h, when compared with air-exposed mice, p ≤ .05. BALF protein, an indicator of air/blood barrier integrity, and W/D were significantly higher, 10- to 12-fold, in phosgene-exposed than in air-exposed mice 4-12 h after exposure, p ≤ .001 and p ≤ .05, respectively. These results indicate that exposure to phosgene causes early bronchoconstriction, a temporal obstructivelike injury pattern, and disruption of mechanical rhythm largely regulated by the progressive production of pulmonary edema on airway flow. Potential therapeutic intervention may include compounds that produce bronchodilation and mechanical ventilation support if warranted.

Effect of Inositol-Trisphosphate on Fluid Transport and Protein Extravasation in the Obstructed Small Bowel

Background: Small-bowel obstruction is characterized by accumulation of fluid in the obstructed intestine. A pronounced inflammation in the obstructed gut wall has been shown to play an important role in the pathogenesis of the profuse fluid losses. α -Trinositol (D-myo-inositol-1,2,6-trisphosphate; PP56) has potent anti-inflammatory as well as antisecretory properties. The effects of α -Trinositol on inflammation and fluid losses in the obstructed small intestine are examined. Methods: A total obstruction of the proximal part of the rat jejunum was induced during 18 h by thread ligation. A small segment of the obstructed intestine, with intact vascular and nervous supply, was placed in a chamber suspended from a force displacement transducer allowing for continuous registration of net fluid transport on a Grass polygraph. Three groups were included. One group ( n = 12) received low-dose α -Trinositol (bolus: 2 mg kg -1 ; IV infusion: 10 mg kg -1 min -1 ), a second group ( n = 10) received high-dose α -Trinositol (bolus: 12 mg kg -1 ; IV infusion: 60 mg kg -1 min -1 ), while a control group ( n = 9) received corresponding volumes of isotonic saline (bolus: 0.5 ml; IV infusions 15 μl min -1 ). Quantitative measurement of extravasated Evans blue albumin in the obstructed jejunum was used as a marker of inflammation. Results: High-dose α -Trinositol induced a significant ( P < 0.001) inhibition of net fluid secretion, while low-dose α -Trinositol had no significant effect versus saline. In contrast, both doses of α -Trinositol induced significant inhibition of EB-albumin leakage ( P < 0.05). Conclusion: High-dose α -Trinositol is a potent inhibitor of fluid secretion in obstructive ileus, most probably involving an anti-inflammatory mechanism.

Biochemical and functional characterisation of eggshell matrix proteins in hens

The eggshell of the hen is a highly ordered, mineralised structure deposited within an acellular milieu – the uterine fluid secreted by the distal oviduct. Spherulitic crystal growth is initiated by deposition of calcium carbonate on aggregates of organic material present on the outer surface of the eggshell membranes. Gel electrophoresis reveals a complex array of proteins in uterine fluid and eggshell extracts. The eggshell matrix proteins can be classified as egg white proteins (lysozyme, ovalbumin, ovotransferrin, clusterin), bone protein (osteopontin), or proteins specific to the uterus and eggshell (ovocleidins-17 and –116; ovocalyxins-32 and –36). Eggshell extracts, uterine fluid and purified fractions are able to modify the morphology of calcite crystals in vitro. In young hens the breaking strength of the eggshell is inversely related to the degree of calcite orientation. Conversely, reduced strength in eggshell from aged hens coincides with a high variability in crystallographic texture. In guinea fowl the exceptional mechanical properties of the eggshell are explained by an increase in the amount of eggshell produced and particular features of the crystallographic texture. These observations suggest that the eggshell matrix influences the process of crystal growth by controlling size, shape and orientation of calcite crystals. This structural control probably contributes in a substantial manner to the mechanical properties of eggshell.

Effects of carbocisteine on airway inflammation and related events in SO2-exposed rats

Airway inflammation leads to secretion of abnormal mucous glycoprotein and ciliary injury. To investigate the possible usefulness of carbocisteine against airway inflammation and events related to it, we conducted a study in SO2-exposed rats of the effects of carbocisteine and ambroxol, as an active control drug, on components of mucous glycoprotein (fucose, sialic acid and protein) in bronchoalveolar lavage fluid (BALF); on infiltration and activation of inflammatory cells in BALF; on tracheal and bronchial-ciliary lesions; and on cAMP levels in tracheal and alveolar tissues. Carbocisteine inhibited or improved all SO2-induced changes tested, and dosages of 125 and 250 mg/kg b.i.d. reduced fucose, sialic acid and protein contents, inflammatory cells (as markers of inflammation), free radicals, and elastase activity in BALF, and suppressed the development of ciliary lesions of the tracheal and bronchial mucosa, while ambroxol (10 mg/kg b.i.d.) showed no such effects. In addition, carbocisteine improved cAMP levels in the tracheal and alveolar tissues. These results indicate that carbocisteine is able to prevent the development of inflammation-related respiratory disease in this rat model, and that this remission of airway inflammation may be associated with carbocisteine-induced normalization of cAMP levels in tracheal and alveolar tissues as well as with its mucoregulant and anti-inflammatory effects. In conclusion, carbocisteine has a unique mucoregulant action and inhibits SO2-induced airway inflammation in a manner different from that of ambroxol.

Capillary Zone Electrophoresis of Proteins in Body Fluids: Comparison of Capillary and Agarose Gel Electrophoresis

The classification of pleural effusions as transudates or exudates often is an early step in their evaluation (1)(2)(3)(4). Exudates but not transudates require additional and often invasive diagnostic procedures. The criteria of Light et al. (5) for classification of pleural effusions have been reported to lead to unwarranted invasive diagnostic procedures in 20–30% of patients with a transudate (1)(3) and to misclassify some exudates as transudates (3)(4). Protein analysis has been proposed to improve classification into transudates or exudates based on the assumption that large proteins are present only in exudates because of the increased capillary permeability (1). Agarose gel electrophoresis (AGE) has been the standard procedure for serum protein fractionation for >30 years, but it is laborious and methods do not agree (6)(7)(8). Capillary zone electrophoresis (CZE) is an attractive alternative (6)(9)(10) and avoids between-protein differences in dye affinity. We evaluated the Beckman Paragon CZETM 2000 system, a multicapillary instrument for automated serum protein electrophoresis (7)(8), for quantitative fractionation of proteins in body fluids. For method comparison, leftover samples from 49 pleural and 11 ascitic fluids from 47 inpatients were used. There were no preset selection criteria regarding age, sex, or type of disease (11); however, only 37 pleural effusions were classified as transudates or exudates after exclusion of patients lacking definitive clinical diagnosis (n = 5), patients with multiple diagnoses (n = 3), or in case of interference by radioopaque agents (n = 4). A serum sample was collected simultaneously with the pleural fluid (n = 30) or within 24 h before (n = 13) or after (n = 17) the puncture. Final diagnosis was retrieved from the patient’s file based on the conclusions reached by …

Pleural Protein Capillary Electrophoresis for the Separation of Transudates and Exudates

Categorization of pleural effusions as transudates or exudates assists diagnostic and therapeutics decisions. To meet the criteria of Light et al. (1) for exudates, an effusion must have at least one of the following: a ratio of pleural fluid (PF) protein to serum protein >0.5, a ratio of PF to serum lactic dehydrogenase (LD) >0.6, and PF LD more than two-thirds the upper limit of normal for serum LD. Numerous studies have examined the diagnostic accuracy of these criteria, which misdiagnose 10–30% of transudates as exudates (2)(3). Recently in this Journal, Chen and Lam (4) reported that qualitative protein zone electrophoresis is more sensitive (100% vs 95%) and specific (50% vs 38%) than the criteria of Light et al. (1) in a study of 51 patient samples (8 transudates and 43 exudates). Moreover, when quantitative analysis was performed, the PF α2-globulin:albumin ratio at the best cutoff point (0.28) showed a sensitivity and specificity of 85% and 80%, respectively. To determine whether protein capillary electrophoresis rather than protein zone electrophoresis meets the accuracy of the criteria of Light et al. (1), we prospectively studied 116 adult patients with pleural effusions over a 1-year period. On the basis of predetermined clinical criteria (2)(3)(4), there were 29 … [↵][1]bAuthor for correspondence. Fax 852-2636-5090; e-mail ching-wanlam{at}cuhk.edu.hk. [1]: #xref-corresp-2-1

Food allergy: what are the issues?

With a growing interest in the development of genetically modified crop plants there is a need for appropriate approaches to safety assessment. Among the issues that have to be addressed is consideration of whether the products of novel genes have the potential to cause allergic sensitization. Resulting from a collaboration between the International Food Biotechnology Council and the International Life Sciences Institute recommendations have been made for a step-wise approach to the assessment of allergenic potential based upon considerations of serological identity, and sequence or structural homology, with known allergens and examination of the stability of the test protein in a simulated gastric fluid. In parallel there has been interest in the development of animal models, which would permit a more direct evaluation of potential allergenic activity. Progress in these areas is reviewed briefly in the context of what is known of food allergy and some of the important issues, which must be addressed in designing safety assessment strategies identified.

The role of nitric oxide in hyperoxic lung injury in premature rats.

To investigate the role of nitric oxide (NO) in hyperoxic lung injury, the 3-day-old preterm rats were randomly assigned to four groups: group I (hyperoxia group), group II (hyperoxia + Nw-nitro-L-arginine methyl ester (L-NAME) group), group III (air group), and group IV (air + L-NAME) group. Group I and II were exposed to > or = 90% O2 for 3 or 7 days. Group II and IV received subcutaneous L-NAMEy on daily basis (20 mg/kg). After 3 day or 7 day exposure, the lung wet weight/dry weight ratio (W/D), total protein and malondialdehyde (MDA) in bronchoalveolar lavage fluid (BALF) and lung pathology were examined in all groups. NO content, expression of endothelial NOS (eNOS) and inducible NOS (iNOS) in lungs were measured in group I and III. Our results showed that after 3 day exposure, group I appeared acute lung injury characterized by the increase of MDA content (P < 0.01) and the presence of hyperaemia, red cell extravasation and inflammatory infiltration; after 7 day exposure, except MDA, total protein and W/D were also increased in comparison with group III (P < 0.01, 0.05), pathological changes were more severe than those after 3 day exposure. After 3 and 7 day exposure, total protein in group II was significantly increased as compared with group I (P < 0.01 for both). The pulmonary acute inflammatory changes were more obvious in group II than in group I. Occasionally, mild hemorrhage was detected in the lungs of group IV. BALF protein content in group IV was higher than that in group III after 7 day exposure (P < 0.01). After 3 and 7 day exposure, NO content in BALF were all significantly elevated in group I as compared with group III (P < 0.01 for all). In the lungs of group I, strong immunostaining for iNOS was observed in airway and alveolar epithelia, inflammatory cells, which were stronger than those in group III. Expression of iNOS in rats after 7 day hyperoxic exposure was stronger than that after 3 day exposure. Shortly after 7 day exposure, stronger immunostaining for eNOS in airway epithelia in group I than that in group III was seen. Our study suggested that treatment with L-NAME worsened acute hyperoxic lung injury in preterm rats and also had a deleterious effect on the rats exposed to air, indicating that endogenous nitric oxide may play a protective role in rats under both physiological and hyperoxic status. Hyperoxia can significantly upregulate the expression of iNOS and eNOS in inflammatory cells, epithelia in the lungs of preterm rats, promote NO generation, which suggests that endogenous NO may mediate the hyperoxic pulmonary damage. Over-stimulation of iNOS may contribute to the pathogenesis of hyperoxic lung injury. NO may have dual roles in pulmonary oxygen toxicity.

IL-5 and TNF-alpha participate in recruitment of eosinophils to intestinal mucosa in ulcerative colitis.

There is an increased influx of activated eosinophils to the intestinal mucosa in active ulcerative colitis, and an increased release of eosinophil-derived proteins, such as ECP, has also been observed. These findings indicate that eosinophils may contribute to tissue damage and intestinal inflammation in this disease. The relative importance of different chemotactic factors and the impact of steroid treatment on their effect in active ulcerative colitis are not known. We measured the eosinophil chemotactic activity in perfusion fluids from 11 patients with ulcerative colitis before and after steroid treatment and from 7 control patients. The effect of neutralizing antibodies to IL-5 and -8, RANTES, eotaxin, MCP-3, TNF-alpha, GM-CSF was investigated. The chemotactic activity was higher in perfusion fluids from patients than from controls (P = 0.0043). Anti-IL-5 (P = 0.005) and -TNF-alpha (P = 0.017) inhibited the activity in perfusion fluids obtained before treatment. Steroid treatment prevented the effect of all antibodies but had no significant effect on the chemotactic activity. The chemotactic activity correlated with the levels of eosinophil granule proteins in the perfusion fluids. In conclusion, in ulcerative colitis, eosinophils are attracted to the intestinal tissue by chemotactic factors, of which IL-5 and TNF-alpha may be the most prominent steroid-sensitive ones. The steroid-insensitive chemotactic activities remain unidentified.

Cellular and Molecular Mechanisms Regulating Silica-Induced Adhesion Molecule Expression in Mice

Exposure of mice to silica particles generates an intense inflammatory response characterized by the influx of neutrophils and monocytes into the alveoli with resulting cell activation. These cell trafficking and effector functions are, in part, mediated by different families of adhesion molecules. One such adhesion protein, intercellular adhesion molecule-1 (ICAM-1), is expressed on a variety of cells including vascular endothelial cells, fibroblasts, types I and II alveolar epithelial cells, some lymphocytes, and monocytes/macrophages. Over the last several years, our laboratory has focused on silica-induced ICAM-1 expression in exposed mice (in vivo) and cells (in vitro) by examining the role and regulation of this adhesion protein on pulmonary cells. We have determined that intratracheal exposure of mice to silica (2 mg per mouse) elicits increased expression of ICAM-1 on pulmonary macrophages and type II epithelial cells within the lung parenchyma, on bronchoalveolar lavage macrophages, and as a soluble protein in lavage fluid. We hypothesize that increased ICAM-1 expression mayparticipate in silica-induced neutrophil influx into the alveoli, as well as in macrophage-derived phlogistic signals responsible for migration of neutrophils. ICAM-1 expression on silica-exposed mouse macrophages is enhanced by reactive oxygen species (ROS) and tumor necrosis factor-alpha (TNF-alpha) and appears to be regulated through specific sequence elements within the ICAM-1 promoter.

Absence of protease-resistant prion protein in the cerebrospinal fluid of Creutzfeldt-Jakob disease.

Creutzfeldt-Jakob disease (CJD), believed to be caused by a protease-resistant isoform of prion protein (PrP(Sc)), usually manifests itself as a clinically distinctive age-related dementia because of its rapid progression, occasionally accompanied by cerebellar ataxia. Recently, a variant CJD (vCJD) has been described, which has prominent early psychiatric symptoms and an earlier age of death. Although cerebrospinal fluid (CSF) is part of the extracellular fluid of the central nervous system (CNS), the bulk of its proteins are derived from the plasma and there is increasing concern about possible transmission of prion disease by blood. As investigation of CSF has played a significant role in the diagnosis and management of several neurological diseases, it was decided to characterize PrP present in the CSF of CJD individuals. Significant variation was observed in the level of PrP in the CSF from both non-CJD and CJD (including vCJD) patients, and the detected PrP forms are protease-sensitive. Using a conformation-dependent immunoassay, it was further demonstrated that the PrP detected in the CSF from CJD patients was broadly similar in conformation to that found in non-CJD patients. Taken together, the results of this study fail to demonstrate any correlation between the presence of protease-resistant PrP isoform (PrP(Sc)) in the CSF and disease manifestation.

An enzyme‐linked immunosorbent assay to quantify 14‐3‐3 proteins in the cerebrospinal fluid of suspected Creutzfeldt‐Jakob disease patients

An enzyme‐linked immunosorbent assay to quantify 14‐3‐3 proteins in the cerebrospinal fluid of suspected Creutzfeldt‐Jakob disease patients

Creutzfeldt-Jakob Disease: Correlation of Clinical Neuroradiological Aspects and Detection of 14.3.3. Protein in Cerebrospinal Fluid

Creutzfeldt-Jakob disease (CJD) is a rare, transmissible illness that usually affects older adults and is characterised by rapidly progressive dementia, ataxia, myoclonus and various other neurological defects. This is now thought to be a chronic infections disease caused by an agent called a prion. The prions differ from viruses because contain little or no nucleic acid and evoke no immune response. CJD is a progressive, inevitably fatal disease with a mean survival time of 6 months to 1 year. Diagnosis is usually made clinically and electroencephalographically but even now, the diagnosis can only be confirmed by brain biopsy. CT scans usually show either no abnormalities (80% of cases) or non specific atrophy. Recently there have been reports of characteristic findings on MR images. Bilateral, symmetrically increased signal intensity was demonstrated in the basal ganglia but in various patients this finding was absent. Recently, diffusion weighted MR images proved to be useful in the evaluation of CJD but these studies must be confirmed with additional patients. The biomedical analysis of cerebrospinal fluid that identifies 14.3.3. protein in a patient with dementia was positive in 96% of cases. The presence of 14.3.3. protein in cerebrospinal fluid may be due to massive neuronal disruption and to the leakage of brain proteins into cerebrospinal fluid. Also if this protein could sometimes be found in patients with acute viral encephalitis, strokes (within one month before testing), subarachnoid haemorrhage or Rett's syndrome, CJD could not reasonably be included in the differential diagnosis of any of these disorders. We describe three cases of CJD. In two patients the typical MRI images of hyperintensity on T2-weighted images of the basal ganglia were observed. In all three patients 14.3.3. protein was found in the cerebrospinal fluid. Therefore, the association of a clinical diagnosis of dementia, MR imaging and a positive test of 14.3.3. protein in the cerebrospinal fluid strongly supports the diagnosis of CJD without the need for cerebral biopsy. Biopsy could be reserved only for doubtful cases.

Quantification and Characterization of Pregnancy-associated Complexes of Angiotensinogen and the Proform of Eosinophil Major Basic Protein in Serum and Amniotic Fluid

The proform of eosinophil major basic protein (ProMBP) exists in serum from pregnant women complexed with a variable fraction of angiotensinogen (Ang). A subfraction further binds complement C3dg in a 2:2:2 complex. The function, physiology, and clinical importance of ProMBP complexes are unknown, and the specific quantification of these complexes has not been possible. We developed an ELISA for the ProMBP/Ang complexes, using a monoclonal antibody against ProMBP for capture and a chicken anti-human Ang antiserum for detection. Calibrators were standardized with WHO IRP 78/610 for pregnancy proteins in the assay range 0.95-15.6 mIU/L. The concentrations of ProMBP/Ang complexes in serum of nonpregnant blood donors (n = 79) were log-normally distributed with a central 95th interval of 985-3655 mIU/L. In pregnancy, mean serum concentrations were increased from week 7, and the concentrations reached term concentrations in week 18. ProMBP/Ang complexes eluted in gel filtration as a broad peak with a molecular mass of approximately 230 kDa. The concentration of ProMBP/Ang/C3dg increased during blood coagulation, suggesting that the ProMBP/Ang/C3dg complex may be a marker of complement activation. ProMBP/Ang complexes are present in serum from nonpregnant persons as well as pregnant women, and the direct assays described here will make it possible to study the biochemistry and the clinical significance of different ProMBP complexes in pathological conditions and pregnancy.

Spectrophotometric and chromatographic determination of sulfanilamides in biological fluids and pharmaceuticals

The conditions for the determination of norsulfazol (I), sulfadimezine (II), ethazole (III), sulfamethoxypyridazine (IV), sulfadimethoxine (V), sodium sulfacyl (VI), sulfalene (VII), sulgin (VIII), and bucarban (IX) as their derivatives of 4,6-dinitrobenzofuroxan using spectrophotometry and thin-layer chromatography were selected. The best results were obtained at 490–500 nm using polar solvents and their aqueous mixtures of pH 6.68 as analytical media. The analytical range for sulfanilamides was 0.5–6.0 Μg/mL. The limits of detection (Μg/mL) were 0.24 for 1,0.28 for II, 0.43 for III, 0.18 for IV, 0.25 for V, 0.54 for VI, 0.54 for VII, 0.17 for VII, and 0.31 for IX. Sulfanilamides were determined in Pharmaceuticals (tablets, ointments, and drops) and biological fluids (protein hydrolyzates, urine, blood serum, and whole blood). A simple rapid method for the determination of the phenotype of biotransformation by the type of acetylation was developed based on the studies of the kinetics of the excretion of II from the human body with urine.

Diuretics and kidney diseases

The main indications for diuretic treatment of renal patients are edema and hypertension. Pharmacokinetics and pharmacodynamics of diuretics are altered in patients with proteinuria and/or impaired renal function. These patients exhibit avid sodium retention. Diuretics are partially inactivated by binding to proteins in tubular fluid. The natriuretic response to diuretics is limited by counter-regulation, specifically increased proximal tubular reabsorption in response to hypovolemia and increased distal tubular sodium reabsorption in response to increased sodium load. At higher serum creatinine values, thiazides are no longer sufficiently effective in monotherapy. The diuretics of first choice are then loop diuretics, potentially in combination with thiazide diuretics. Potassium-sparing diuretics are contraindicated. The most important side effects of diuretics are hypovolemia with orthostatic hypotension, hypokalemia, metabolic alkalosis, increase of creatinine concentration and (rarely) hyponatremia. Diuretic treatment should be accompanied by reduction of dietary sodium intake. Important points are selection of an adequate dose (in problematic cases dose-finding via urine sodium measurements) and selection of proper dosing intervals. If cases do not respond to loop diuretic monotherapy, combination with thiazide diuretics or intravenous administration of loop diuretics should be considered.

Phosphorylated tau protein in cerebrospinal fluid is elevated in Alzheimer's disease and other tauopathies

Phosphorylated tau protein in cerebrospinal fluid is elevated in Alzheimer's disease and other tauopathies

Mesenteric Lymph Duct Ligation Provides Long Term Protection Against Hemorrhagic Shock-induced Lung Injury

Recently we have shown that ligation of the main mesenteric lymph (MLN) duct prior to an episode of hemorrhagic shock (HS) prevents shock-induced lung injury. Yet, ligation or diversion of intestinal lymph immediately prior to injury is not clinically feasible. Diversion of intestinally derived lymph after injury to protect against secondary insults is possible, but it is not known how long the protective effects of lymph ligation would last. Thus, we tested whether ligation of the MLN duct seven days prior to HS would still be protective. Male Sprague-Dawley rats were subjected to laparotomy with or without MLN duct ligation. Seven days later, half of the sham and actual MLN duct ligated animals randomly were selected to undergo HS (30 mmHG for 90 min). The other half of the animals was subjected to sham shock. Lung permeability, pulmonary myeloperoxidase (MPO) activity, and bronchoalveolar fluid (BALF) protein content were used to determine lung injury. Lymphatic division 7 days prior to HS continued to prevent shock induced lung injury as assessed by a lower Evans Blue dye concentration, BALF protein and MPO activity. In addition, there was no evidence of Patent Blue dye in the previously ligated MLN duct. Since ligation of the main mesenteric lymphatic duct continues to protect against shock-induced lung injury 1 week after duct ligation, it is feasible that lymphatic ligation performed after an injury remains protective against certain secondary insults for at least 1 week.

Evaluation of cellular and biochemical parameters of blood and peritoneal fluid following exploratory laparotomy in the goat.

To evaluate the effects of exploratory laparotomy on cellular and biochemical parameters of blood and peritoneal fluid, an experiment was conducted using 10 Iranian cross-bred male goats. Approximately 10 ml of blood and 1-1.5 ml of peritoneal fluid were collected from all animals prior to operation for estimation of control values. Exploratory laparotomy was performed under local analgesia. Blood and peritoneal fluid samples were collected at 24, 48, 72 and 96 h after exploratory laparotomy. The results revealed that after exploratory laparotomy, the number of white blood cells and the percentage and absolute number of neutrophils and band neutrophils significantly increased (P < 0.05). However, the percentage of lymphocytes decreased significantly (P < 0.05). The concentrations of blood urea nitrogen significantly increased (P < 0.05). Furthermore, following the operation, the percentage and absolute number of neutrophils in the peritoneal fluid significantly increased (P < 0.05). In contrast, the percentage of lymphocytes in the peritoneal fluid decreased significantly (P < 0.05). The concentration of protein in the peritoneal fluid increased significantly (P < 0.05).