Neutrophil Leukocyte Infiltration Research Articles

Pathology Quiz Case 1

A 23-YEAR-OLD WHITE WOMAN PRESENTED with a 2-month history of a pyoderma gangrenosum–like ulcer in the right postauricular region. The cutaneous lesion had initially started as a pustule that was associated with local pain, subfebrile temperature, night sweats, and spontaneous perforation with a whitish discharge 2 weeks after the disease onset. Necrectomy was performed, and antibiotics (clindamycin, amoxicillin and clavulanate potassium [co-amoxiclav], and azithromycin) were administered intravenously in an outpatient clinic, without success. Two months after the first symptoms appeared, the patientwasadmitted toourclinic.Shehada4.5-cm-diameter, painful, necrotic area in the right postauricular and subauricular regions. The lesion was covered with granulation tissue and mucous discharge. In the left postauricular region, there was an indurated purple pustule, 2 cm in diameter, that was solid and painful to palpation. A smaller pustule was seen in the medial part of the left scapula. The patient also had painful episcleritis and uveitis of both eyes as well as granulation tissue in the septal and conchal mucousmembranes.Antibiotic therapy(piperacillinandgentamicinsulfate)wasadministeredintravenously,andasecond necrectomyoftherightpostauricularandsubauricularareas was performed on day 5 of admission. A huge ulcer penetrating deeply into the right parotid gland, almost to the facial nerve, was found on removal of the necrotic tissue (Figure 1). The tissue was sent for cytologic, microbiologic, andhistopathologic analysis.Cultureswerenegative for aerobic and anaerobic microorganisms and fungi. Cytologicexaminationshowedgranulomatous inflammation. Thehistopathologyreport revealed fibrinoidnecrosisof the vascular wall; macrophage, lymphocyte, and neutrophilic leukocyteinfiltration;andformationofgranuloma(Figure2 and Figure 3). What is your diagnosis?

Effect of the flavonoid quercetin on inflammation and lipid peroxidation induced by Helicobacter pylori in gastric mucosa of guinea pig

Helicobacter pylori infection induces an inflammatory response in the gastric mucosa. Activation of polymorphonuclear leukocytes can produce oxidative damage to gastric tissue through intermediary radicals of oxygen and nitrogen. Vegetable extracts containing polyphenols of the flavonoid family have antibacterial activity, and the flavonoid quercetin possesses anti-H. pylori activity in vitro. The aim of this study was to analyze the effect of oral administration of pure quercetin on inflammation and lipid peroxidation induced by H. pylori in the gastric mucosa of the guinea pig. Sixty days after oral infection with H. pylori guinea pigs received 200 mg/kg of quercetin daily by mouth for 15 days. The infiltration index of inflammatory cells and bacterial density in both the pyloric antrum and corpus were histologically determined by myeloperoxidase histochemistry, hematoxylin-eosin, and modified Giemsa stains. The lipid hydroperoxide content was assessed by the orange xylenol spectrophotometric method. Quercetin significantly reduced the infiltration index of mononuclear cell and bacterial colonization in the pyloric antrum and corpus. In the antrum of infected quercetin-treated animals, a significant diminution of neutrophil leukocyte infiltration was observed compared with the infected nonquercetin-treated animals. In the antrum, the lipid hydroperoxide concentration was significantly decreased in infected animals treated with quercetin, whereas in the corpus no significant differences were observed. Our results indicate that in vivo oral quercetin administration decreases H. pylori infection in the gastric mucosa and reduces both the inflammatory response and lipid peroxidation.

Effects of sulfasalazine on biopsy mucosal pathologies and histological grading of patients with active ulcerative colitis

To investigate the mechanisms of sulfasalazine (SASP) in the treatment of ulcerative colitis (UC). Changes of pathological signs and histological grading of 106 patients with active UC were observed before and after the treatment with SASP, 1 g, thrice daily for 6 wk. The effect of SASP on the vasculitis in lamina propria was 48.2% and 17.4% in the mild active UC (P<0.001) and 68% and 26.7% in the moderate active UC (P<0.001) before and after treatment. Fibroid necrosis of vessel wall was found in one case of mild UC and two cases of moderate UC before treatment and was not found after treatment. No thrombosis was found in mild UC before and after treatment, while thrombosis was found in one case of moderate UC before treatment. The effect on mucosal glandular abnormality was 30.4% and 13.0% in mild UC (P<0.05), and 42% and 40% in moderate UC (P>0.05) before and after treatment. The rate of eosinophil infiltration was 98.2% and 80.4% in mild UC (P<0.01), and 100% and 91.1% in moderate UC (P<0.05) before and after treatment. The effect on crypt abscess was 21.4% and 4.4% in mild UC (P<0.05), and 48% and 13.3% in moderate UC (P<0.001) before and after treatment. The effect on mucosal pathohistological grading was 2.00+/-0.84 and 0.91+/-0.46 in mild UC (P<0.001), and 2.49+/-0.84 and 1.31+/-0.75 in moderate UC (P<0.001) before and after treatment. SASP can improve small vessel lesions and crypt abscesses and reduce neutrophilic and eosinophilic leukocyte infiltration in inflammatory mucosa of UC.

The role of the glucocorticoid-dependent mechanism in the progression of sodium taurocholate-induced acute pancreatitis in the rat.

The effects of glucocorticoids on acute pancreatitis (AP) have remained contradictory. The aim of this study was to investigate the time courses of the effects of the exogenous glucocorticoid agonists dexamethasone (DEX) and hydrocortisone (HYD) and a glucocorticoid antagonist (RU-38486) and to characterize the local and systemic responses in AP in rats. The glucocorticoid antagonist and agonists were administered just before AP induction. Serum amylase activity determinations, IL-6 bioassays, pancreatic weight/body weight ratio measurements, and survival analysis were performed. Liver and lung injuries were assessed via neutrophil leukocyte infiltration in myeloperoxidase (MPO) assays, tissue adenosine triphosphate (ATP) level determinations, and histology. In the glucocorticoid agonist groups, the survival rate increased, while the serum amylase level, the IL-6 activity, and the pancreatic weight/body weight ratio decreased significantly as compared with the control and RU-treated groups. AP resulted in significant decreases in tissue ATP levels in both the liver and the lung. In the DEX- or HYD-treated groups, the liver ATP levels were significantly elevated, while both the liver and the lung MPO levels were attenuated as compared with the AP and RU-treated groups. These results suggest that glucocorticoids may play important roles in mitigating the progression of the inflammatory reaction during the early phases of AP.

The Pathological Change of Zedoary Turmeric Oil Gelatin Microspheres Administered Via Hepatic Artery

目的 观察莪术油明胶微球栓塞Beagle犬肝动脉后的病理改变,为临床实验提供依据.方法 Beagle犬随机分为生理盐水组(10ml/kg)、空白微球组(15mg/kg)、莪术油微球低剂量组(7.5mg/kg)和莪术油微球高剂量组(15mg/kg),每4周经肝动脉给药1次,连续给药3次,停药后观察4周.观察肝病理组织学改变.结果明胶微球肝动脉栓塞可致肝组织不可逆性损伤,高剂量组广泛全小叶坏死,低剂量组灶性坏死,空白微球组改变类似高剂量组,生理盐水组未见肝组织损伤.结论莪术油微球重复经Beagle犬肝动脉给药,可因肝动脉栓塞造成肝脏不可逆性缺血坏死。

Additive effects of phosphodiesterase-4 inhibition on effects of rSP-C surfactant.

We have tested the effects of combined treatment with a recombinant surfactant protein C based surfactant (rSP-C surfactant) containing a phosphodiesterase-4 (PDE-4) inhibitor, roflumilast, in a lung lavage model of acute lung injury. The following groups were tested: (1) controls receiving sham exposure; (2) PDE-4 inhibitor (6.0 mg/kg body weight, intratracheally) alone; (3, 4) rSP-C surfactant (25 and 100 mg phospholipids [PL] per kg body weight) alone; and (5, 6 ) treatment with rSP-C surfactant (25 and 100 mg PL per kg body weight) combined with the PDE-4 inhibitor at a dose of 6.0 mg/kg body weight. The different groups were compared with respect to improving oxygenation and histopathologic changes, e.g., hyaline membrane (HM) formation. Both doses of rSP-C surfactant improved oxygenation while even this high dose of the PDE-4 inhibitor alone did not influence oxygenation compared with untreated control animals. Addition of the PDE-4 inhibitor led to improved oxygenation based on both doses of rSP-C surfactant. The PDE-4 inhibitor alone prevented further HM formation and infiltration of neutrophil leukocytes. The rSP-C surfactant was able to prevent further HM formation. Based on both doses of rSP-C surfactant, addition of the PDE-4 inhibitor showed additional effects on oxygenation and inhibition of HM formation. The effects of combined treatment were superior to each treatment alone, leading to the conclusion that a rSP-C surfactant containing a PDE-4 inhibitor may act synergistically in this animal model of acute lung injury. We conclude that combined treatment with rSP-C surfactant and a PDE-4 inhibitor may be an effective treatment for patients with acute lung injury.

Dexamethasone enhances the activity of rSP-C surfactant but not of Exosurf in a rat model of the acute lung injury

The possible enhancement of surfactant activity by pretreatment with a glucocorticosteroid (dexamethasone) was investigated in a rat lung lavage model of acute lung injury. Animals received a dose of dexamethasone (10 mg/kg i.p.) prior to the protein-free surfactant preparation Exosurf (pure phospholipids containing surfactant, Wellcome GmbH, Burgwedel, Germany) and a rSP-C based surfactant, respectively. Both surfactants were intratracheally instilled at doses of 25 and 100 mg phospholipids per kg body weight and were compared with pretreatment with dexamethasone at each dose level. These groups were also compared with untreated controls and to pretreatment with dexamethasone alone with respect to improvements in oxygenation, to inhibition of infiltration of polymorphonuclear neutrophil leukocytes (PMNL) and to influence formation of hyaline membranes. Dexamethasone alone had no influence on the reduced PaO 2 but reduced the infiltration of PMNL and the formation of hyaline membranes. Dexamethasone improved the oxygenation at both doses of rSP-C surfactant. At the low dose of rSP-C surfactant there were additional effects detectable with regard to histopathologic improvements. In contrast, dexamethasone had no additional effect on oxygenation and formation of hyaline membranes when combined with Exosurf. Only the infiltration of PMNL was decreased by combined treatment with dexamethasone and Exosurf. The effect was comparable to that of pretreatment with dexamethasone alone. In this animal model, pretreatment with dexamethasone showed additional effects on rSP-C surfactant that were superior to each treatment alone. From the comparison of rSP-C surfactant with the synthetic surfactant preparation Exosurf, we conclude that the activity of Exosurf cannot be improved substantially by additional pretreatment with drugs like glucocorticosteroids.

A rat model of acute respiratory distress syndrome (ARDS) Part 2, influence of lavage volume, lavage repetition, and therapeutic treatment with rSP-C surfactant

The influence of lavage volume, and lavage repetition with physiological saline solution (groups 1–3: 3×4, 4×4, 5×4, groups 7–9: 3×8, 5×8, 7×8, mL per animal) was studied in a rat lung lavage model of the acute respiratory distress syndrome (ARDS). Anesthetized and tracheotomized rats (12 rats/group) were pressure-controlled ventilated with 100% oxygen at a respiratory rate of 30 breaths/min, inspiration: expiration ratio of 1:2, peak inspiratory pressure of 28 cm H 2O at positive end-expiratory pressure of 8 cm H 2O during the whole experimental period. To investigate the influence of therapeutic treatment, a recombinant surfactant protein C (rSP-C) containing surfactant was used. Therefore, rats which received a lavage of 4×4 mL per animal (groups 4 to 6) or 7×8 mL per animal (groups 10–12) were treated intratracheally with surfactant doses of 12.5, 25, or 100 mg phospholipids (PL) per kg body weight (bw). In all groups, partial arterial oxygen pressures (PaO 2, mm Hg) and partial arterial carbon dioxide pressures (PaCO 2, mm Hg) were determined 30 min before, directly after, and 5, 30, 60, 90, 120, 150, 180, and 210 min after the last lavage. Additionally, animals were euthanized 210 min after the last lavage for semiquantitative histopathological grading of coded lung slides. Grading was performed with respect to the severity of hyaline membrane formation (HM), margination and infiltration of polymorphonuclear neutrophil leukocytes (PMNL) into the lung alveoli and interstitial and intraalveolar edema (E). The intrapulmonary distribution of intratracheally applied rSP-C was estimated in selected lung slides stained with polyclonal anti-rSP-C antibody and was compared to unlavaged control rats and unlavaged rats which received 100 mg/kg bw rSP-C. The repetitive lavage depleted the lung from its natural surfactant resources leading to a pathophysiological cascade similar to that of the acute respiratory distress syndrome. PaO 2 levels and HM formation showed a lavage-induced decrease. Both changes were significantly dependent on the repetition and volume of the lavage; however, the parameters PMNL and E did not show such a dependence. Treatment with rSP-C surfactant significantly improved oxygenation and reduced HM-formation in a dose-dependent manner independent from the lavage volume. All doses of rSP-C surfactant showed no clear influence on the parameters PMNL and E independently from the lavage volume. In lavaged rat lungs (ARDS-model), the exogenously applied rSP-C was distributed homogeneously along the alveolar lining. Unlavaged rats that received a similar dose of rSP-C showed a marked inhomogeneous extracellular distribution, mainly associated with larger bronchi, while the type II pneumocytes were stained positively in unlavaged control and unlavaged rSP-C treated rats. Conclusion: This model mimics very closely the wide spectrum of the clinical situation of human acute lung injury (ALI) because the variation of lavage volume and repetition lead to reproducible different severity grades and states of ALI. The significant reduction of pathognomic changes due to treatment with rSP-C surfactant showed that this is a useful model to estimate the influence of therapeutic concepts in ALI and ARDS.

Relationship between endomyometritis and the duration of premature membrane rupture.

In this study, correlation between duration of premature rupture of the membranes (PROM) and the degree of membrane, decidua, and myometrium infections were histologically investigated. The study was carried on 61 women with term pregnancy (30 with PROM, 31 control group). During cesarean section, a full-thickness biopsy specimen of the uterine wall from serosa to decidua was obtained with scissors. Chorioamnionitis was histopathologically identified by neutrophilic leukocyte (NL) infiltration of the membranes. Endometritis or myometritis were diagnosed when the acute inflammatory process extended to the decidua or the myometrium successively. It was found that the presence and depth of NL infiltration significantly correlates with the interval between rupture of membranes and termination of pregnancy.

A rat model of acute respiratory distress syndrome (ARDS): part 1. time dependency of histological and pathological changes

The time course of histopathological changes in a rat lung lavage model of the acute respiratory distress syndrome (ARDS) was analyzed by sacrificing animals 10, 30, 60, 180, and 210 min after the last lung parenchyma lavage which was performed with physiological saline solution. This lavage depleted the lung from its natural surfactant resources leading into a pathophysiological cascade similar to that of the acute respiratory distress syndrome. Tracheotomized rats (12 animals per time point) were pressure-controlled ventilated (Siemens Servo Ventilator 900C) with 100% oxygen at a respiratory rate of 30 breaths/min, inspiration–expiration ratio of 1:2, peak inspiratory pressure of 28 cm H 2O at positive end-expiratory pressure (PEEP) of 8 cm H 2O. During the whole experimental period, the ventilation was not changed. Blood gases (partial arterial oxygen pressures [PaO 2, mmHg] and partial arterial carbon dioxide pressures [PaCO 2, mmHg]) were estimated before, directly after, and 10, 30, 60, 90, 120, 150, 180, and 210 min after the last lavage. For grading lung lavage-induced histopathological changes associated with the time-dependent development of ARDS, slides were coded and evaluated without any knowledge of the sacrifice time. A semiquantitative grading was performed with respect to the severity of the following parameters: hyaline membrane formation (HM), interstitial and intraalveolar edema edema (E), and margination and infiltration of polymorphonuclear neutrophil leukocytes (PMNL) into the lung alveoli. The severity of these parameters showed a time-dependent increase after the last lavage. This was accompanied by a time-dependent decrease in partial arterial oxygen pressure (PaO 2) values during the early postlavage period (up to 30 min). Thereafter, PaO 2 levels remained fairly stable. The severity of intraalveolar and/or perivascular hemorrhages within the lung was not time dependent. The rat lavage model shows similarities to the pathophysiological sequelae occuring during the acute phase of the acute respiratory distress syndrome in humans. Most of the characteristic pathognomic histological changes seen in humans can be observed in this lung lavage model. This ARDS model is brief and easy in its experimental design, showed a good and homogeneous reproducibility of pathophysiological and histopathological parameters, and is therefore a useful model to estimate the influence of therapeutic pharmacological treatments of ARDS.

Long-term effects of repeated injection sclerotherapy on esophageal motility and mucosa.

Endoscopic sclerotherapy (ST), widely used as treatment of bleeding esophageal varices, might cause motility disturbances of the esophagus as well as mucosal damage. We performed this study to evaluate the long-term effects of repeated sclerotherapy on esophageal motility and mucosa. Ten patients with liver cirrhosis and bleeding esophageal varices treated with repeated ST were evaluated after the last ST, median 52 months, by esophageal manometry and gastroscopy where forceps biopsies were taken. We found a significant difference in the distal esophageal sphincter intraabdominal length. The distal esophageal sphincter pressure was somewhat lower in the ST group although the difference did not reach statistical significance. There was infiltration of neutrophil leukocytes in biopsies from four patients and normal findings in the rest. Long-term follow-up evaluation showed statistically longer distal esophageal intraabdominal length in the ST group. No mucosal alterations were found at the histopathological investigation.

Studies on coccidiosis in guinea pigs. 1. Clinico-pathological observation

Clinico-pathological and parasitological studies have been performed on spontaneous and experimental coccidiosis in guinea pigs. Among 11,244 Hartley guinea pigs purchased from suppliers during 1968, 410 (3.6%) of the animals had diarrhea due to coccidiosis. The incidence rate was high in the spring and fall with a mortality rate of 14.4 per cent. A particularly high number of fatal cases were found in the spring. In experimentally induced coccidiosis, clinical signs observed were diarrhea, dehydration, weight loss and death. The diarrhea developed in all animals on the eleventh day after infection and continued for one to five days. Food and water intakes were markedly reduced after the appearance of diarrhea, followed by anorexia and dehydration. Correlating with the appearance of diarrhea was a striking drop in body weight of the guinea pigs. Death usually occurred on the third to fifth day after the onset of diarrhea. The mortality rate was 30 per cent. The major macroscopic findings were characterized by a markedly thickened wall from the ascending to the descending colon and gelatinous edema of the mesenterium of the spiral of the ascending colon. Histologically, there was marked hyperplasia of the mucosal epithelium in the colon and numerous coccidia at different stages of development within the mucus membrane. In the advanced stages of the disease, there was degeneration and desquamation of the epithelia, marked edematous change and infiltration of neutrophil leukocytes and lymphocytes into the lamina propria and submucosa, many oocysts in the lumen of the intestine and in the intestinal glands.(ABSTRACT TRUNCATED AT 250 WORDS)

Experimental candidiasis in the hamster cheek pouch

Sixty-four adult male BIO 87.20 hamsters were divided into four equal groups. Animals in three groups had 1 ml of a thick aqueous suspension of either Candida albicans (ATCC 10261), Candida albicans (UO1) or Candida tropicalis (3100, Puna Culture Collection) placed in each of their cheek pouches. The fourth group were controls. Four animals from each group were killed 1, 2, 4 and 6 weeks following treatment. Specimens were removed from all left-cheek pouches and processed routinely for light microscopy. All animals treated with strain UO1 and 35 per cent of animals treated with either 10261 or 3100 strains exhibited changes. At 1 week, the epithelium had localized areas of neutrophilic leukocyte infiltration and some small discrete micro-abscesses. At 2 weeks, most micro-abscesses were larger. At 4 weeks, most micro-abscesses involved the more superficial epithelial layers. At 6 weeks, there were only a few superficial micro-abscesses. At 4 and 6 weeks, there were areas of thickened, often parakeratinized, stratum corneum. The connective tissue adjacent to the inflamed epithelium was infiltrated by varying numbers and types of chronic inflammatory cell. Hyphal invasion of the epithelium was not found. Thus the hamster cheek pouch is a suitable site for the study of experimental candidiasis. The occurrence of both an acute and chronic inflammatory response does not support the suggestion that the pouch is an immunologically-privileged site.

Microscopic characterization of ocular damage produced by Pseudomonas aeruginosa toxin A.

The ocular damage in young adult mice produced by purified Pseudomonas aeruginosa exotoxin A was microscopically characterized at 1 and 5 h and at 1, 3, 5, 7, 10, 14, and 21 days after toxin A challenge, using light, transmission, and scanning electron microscopic techniques. Similarly to previously described infection with viable organisms, toxin A killed both epithelial and endothelial cells and induced stromal cell swelling within 5 to 24 h after application onto the nonpenetrating wounded corneal surface. Other toxin-induced damage similar to the damage produced by infection with the viable bacteria was production of electron-dense particles within the corneal stroma, dispersal of undamaged collagen fibrils, and apparent loss of stromal proteoglycan ground substance. Toxin A damage differed from infection with the viable bacteria in essentially two ways. First, more purulent exudate and more polymorphonuclear neutrophilic leukocyte (PMN) infiltration of the corneal stroma were produced by infection with the viable organisms than by the toxin. Additionally, PMN did not appear within the toxin-treated corneas until 3 days after treatment, whereas in corneas infected with the viable organisms, PMN were numerous by 18 h. Secondly, toxin A produced cataract of the ocular lens, whereas infection with the viable organisms did not.

Reaction of rat connective tissue to unfixed and formaldehyde-fixed autogenous implants enclosed in tubes

Fixed and unfixed samples of rat connective tissue were enclosed in polyethylene tubes and reimplanted subcutaneously in the animal from which they originated. Histologic studies of the implants indicated that sterile necrotic autogenous tissue elicited a milder response, characterized by less infiltration of polymorphonuclear neutrophil leukocytes (PMN cells), than autogenous tissue fixed in formaldehyde, and was resorbed and replaced by granulation tissue at about the same rate as fixed autogenous tissue that had been washed to remove formaldehyde. Unwashed fixed autogenous tissue fixed in formaldehyde caused necrosis of surrounding tissue and was resorbed at a slower rate than the other implants. The fixation in formaldehyde of autogenous connective tissue was reversible and resulted in complete degeneration and necrosis of the fixed tissue by 15 weeks. Fixed and unfixed samples of rat connective tissue were enclosed in polyethylene tubes and reimplanted subcutaneously in the animal from which they originated. Histologic studies of the implants indicated that sterile necrotic autogenous tissue elicited a milder response, characterized by less infiltration of polymorphonuclear neutrophil leukocytes (PMN cells), than autogenous tissue fixed in formaldehyde, and was resorbed and replaced by granulation tissue at about the same rate as fixed autogenous tissue that had been washed to remove formaldehyde. Unwashed fixed autogenous tissue fixed in formaldehyde caused necrosis of surrounding tissue and was resorbed at a slower rate than the other implants. The fixation in formaldehyde of autogenous connective tissue was reversible and resulted in complete degeneration and necrosis of the fixed tissue by 15 weeks.

Chemical Injury of the Spinal Cord of the Rabbit after Intracisternal Injection of Gentamicin

A single intracisternal injection of 0.4 ml of 1.25 and 2.5 percent gentamicin sulfate with preservative to healthy adult rabbits caused acute respiratory paralysis and severe seizure activity initially and paralysis of the limbs subsequently. In the white matter of the upper cervical spinal cord, multiple, minute, disseminated, spongy lesions were observed. They consisted of lysis of axis cylinders, edematous dilatation of myelin sheaths, and loss of astroglia and interfascicular oligodendroglia. Axonal end-bulbs formed at the periphery of the lesions. Clinically and morphologically, 0.025 and 0.25 percent gentamicin sulfate solution did not produce myelopathy. The spinal lesions were distributed differently from those of other chemical myelopathies in that they developed in the deeper white matter with sparing of marginal myelinated fibers. Circumscribed high concentrations and gentamicin, and vulnerability of myelinated axis cylinders and interfascicular oligondendroglia to gentamicin may be the main factors causing these lesions. When gentamicin sulfate without preservative was injected, neutrophil leukocyte infiltration occurred actively in the spongy lesions. In the cervical spinal ganglia some nerve cells underwent cytoplasmic vacuolation. In control animals a single intracisternal injection of saline or preservative did not result in the production of these lesions.