Modulation of monocyte chemokine production and nuclear factor kappa B activity by oxidants.

Reactive oxygen species can directly damage tissue. In this setting, amplification of tissue damage also occurs through infiltration of inflammatory cells either acutely or chronically. Several recent studies suggest that reactive oxygen species stimulate production of certain chemokines, which are potent chemoattractants for inflammatory cells. In the present study, we examined whether oxidants, generated by the combination of xanthine and xanthine oxidase (X/XO), alter chemokine production by monocytes and U937 cells. Our findings demonstrate that X/XO stimulates monocytes, but not U937 cells, to produce increased amounts of interleukin-8 (IL-8) and monocyte chemoattractant protein. This effect is attenuated by pretreatment with dimethylsulfoxide (DMSO), a scavenger of hydroxyl radicals, but is not affected by superoxide dismutase or catalase. In contrast, X/XO-induced cytotoxicity, evidenced by lactate dehydrogenase release, is mediated primarily by hydrogen peroxide, as catalase reverses this effect. Finally, exposure to X/XO causes an increase in nuclear factor kappa B (NF-kappaB), and this effect is attenuated by DMSO. These studies suggest that reactive oxygen species can induce production of molecules that amplify inflammation through attraction of inflammatory cells. It appears the hydroxyl radical is the principal oxidant species involved in stimulation of chemokine production.

Attenuation of interleukin-8 production by inhibiting nuclear factor-kappaB translocation using decoy oligonucleotides.

Interleukin-8 (IL-8), a monocyte-derived neutrophil chemoattractant factor, is a polymorphonuclear neutrophil chemotaxin that is involved in a number of inflammatory disorders. Transcription of the IL-8 gene is controlled by regulatory proteins, including nuclear factor-kappaB (NF-kappaB), a family of proteins that is important in the transcriptional control of a number of genes. When cells are activated, NF-kappaB translocates from the cytoplasm to the nucleus, where it activates transcription by binding to a specific sequence within the 5' untranslated region of the gene. During translocation, NF-kappaB is potentially susceptible to diversion by oligonucleotides that contain the binding sequence for this protein. In the current study, we produced phosphorothioate-modified oligonucleotides containing the specific DNA sequence that NF-kappaB binds within the IL-8 gene. We then investigated the effects of transfection of monocytes with these oligonucleotides on interleukin-1beta (IL-1beta)-stimulated IL-8 production, IL-8 mRNA expression, and NF-kappaB binding activity. We found that transfection with these oligonucleotides significantly inhibited monocyte IL-8 production. A single-stranded oligonucleotide with two copies of the NF-kappaB-binding sequence was the most potent of those tested. This single-stranded oligonucleotide also inhibited IL-1beta-induced translocation of NF-kappaB to the nucleus and reduced IL-8 mRNA expression. These studies demonstrated that monocyte production of IL-8 can be attenuated using a single-stranded oligonucleotide that binds a transcriptional activating protein before it translocates to the cell nucleus. This approach ultimately may be useful in the control of inflammation involved in a number of diseases.

Calcium-PS-dependent protein kinase C and surfactant protein A in isolated fetal rabbit type II alveolar cells and surfactant-related material.

The fetal lung secretes significant quantities of surfactant during late gestation to prepare for initiation of respiration at birth. However, the mechanism by which this occurs has not been determined. Since Ca2+-phosphatidylserine (PS)-dependent protein kinase C has been implicated in surfactant secretion in adult lung, the present study was done to determine whether this enzyme is also involved in the initiation of surfactant release from fetal type II cells. Type II cells isolated from gestational day-24 fetal rabbits were used. Cells were prelabelled with [32P] and [3H]choline and exposed to 4beta phorbol ester (10(-5) M) for 2 h. Secretion product and subcellular fractions were isolated by removing the culture medium, mixing with homogenate from adult rabbit lung, and subfractionating by centrifugation on a sucrose gradient. Samples of secretion product were also prepared for electron microscopy. Ca2+-PS-dependent protein kinase C was also assayed in some samples, and an add-back technique was used to determine whether enzyme activity in the intracellularly stored surfactant fraction was due to contamination. The results showed that material released by fetal type II cells after exposure to phorbol ester coprecipitated with adult rabbit lung lamellar bodies and microsomes. Morphologically, a range of forms, including lamellar-body-like structures, was detected. The released material originated largely from the lamellar body compartment of the fetal type II cells and displayed immunoreactivity with antibody to surfactant protein A (SP-A) at 35 and 70 kDa apparent molecular mass. Assay of protein kinase C in fetal type II cells showed that exposure to conditioned medium, which induces differentiation, increased activity. Incubation with phorbol ester induced translocation of activity to the microsomal fraction. Add-back assays suggested that protein kinase C activation by treatment with phorbol ester induced translocation of enzyme activity to the lamellar body fraction; none was detected prior to treatment. These results support a role for Ca2+-PS-dependent protein kinase C in initiation of surfactant release by interaction with the developing lamellar body compartment in fetal type II cells.

Correlation of plasma color index with serum bilirubin in neonatal hyperbilirubinemia.

In this study, a simple method was evaluated for serum bilirubin estimation in neonates. The color of plasma was compared with various dilutions of potassium dichromate. Readings were then compared with serum bilirubin levels obtained by diazo reaction. The coefficient of correlation (r = 0.7947) between the two was statistically significant. It is concluded that the color index provides reliable estimates of serum bilirubin levels.

Myelolipoma of adrenal gland in association with occult malignancy--a case report.

Myelolipoma is an unusual lesion of adrenal gland in which ectopic marrow tissue is present. This case is being present because of its rarity.

Study of nucleolar organizer regions (NORs) in lesions of gallbladder.

Tissue sections from 100 specimens of gallbladder including 40 cases of chronic cholecystitis, 30 cases of epithelial hyperplasia and 30 cases of well, moderately and poorly differentiated adenocarcinoma of gallbladder were studied for argyrophillic nucleolar organizer regions (AgNORs). The arithmetic mean +/- S.D. were calculated for each category. This showed NOR counts of chronic cholecystitis--(1.89+/-0.96), epithelial hyperplasia--(3.99+/-1.03), well differentiated adenocarcinoma (WDCA)--(7.04+/-1.34), moderately differentiated adenocarcinoma (MDCA)--(7.52+/-0.97) and poorly differentiated adenocarcinoma (PDCA)--(8.37+/-1.27). The study revealed increasing number of AgNORS from cases of chronic cholecystitis to epithelial hyperplasia to carcinoma gallbladder, but a considerable overlap in AgNOR counts of individual cases was observed suggesting that though AgNOR count cannot act as a specific diagnostic parameter for diagnosis of early carcinoma & dysplasia in isolated cases, they may prove to be a good adjunct to presently available imaging techniques and cytology.

Obesity and outcomes in patients hospitalized with pneumonia.

Studies suggest obesity is paradoxically associated with better outcomes for patients with pneumonia. Therefore, we examined the impact of obesity on short-term mortality in patients hospitalized with pneumonia. For 2 years clinical and radiographic data were prospectively collected on all consecutive adults admitted with pneumonia to six hospitals in Edmonton, Alberta, Canada. We identified 907 patients who also had body mass index (BMI, kg/m(2)) collected and categorized them as underweight (BMI < 18.5), normal (18.5 to <25), overweight (25 to <30) and obese (>30). Overall, 65% were >65 years, 52% were female, and 15% reported recent weight loss. Eighty-four (9%) were underweight, 358 (39%) normal, 228 (25%) overweight, and 237 (26%) obese. Two-thirds had severe pneumonia (63% PSI Class IV/V) and 79 (9%) patients died. In-hospital mortality was greatest among those that were underweight (12 [14%]) compared with normal (36 [10%]), overweight (21 [9%]) or obese (10 [4%], p <0.001 for trend). Compared with those of normal weight, obese patients had significantly lower rates of in-hospital mortality in multivariable logistic regression analyses: adjusted odds ratio (OR), 0.46; 95% CI, 0.22-0.97; p 0.04. However, compared with patients with normal weight, neither underweight (adjusted OR, 1.13; 95% CI, 0.54-2.4; p 0.7) nor overweight (adjusted OR, 0.94; 95% CI, 0.52-1.69; p 0.8) were associated with in-hospital mortality. In conclusion, in patients hospitalized with pneumonia, obesity was independently associated with lower short-term mortality, while neither being underweight nor overweight were. This suggests a protective influence of BMIs > 30 kg/m(2) that requires better mechanistic understanding.

The Immunohistochemical Expression of the Oestrogen Receptor (ER), HER-2/NEU and Cytokeratin 8/18 and 5/6 in Invasive Breast Carcinoma.

INTRODUCTION: Carcinoma is the most common malignancy of the breast and breast cancer is the most common non-skin malignancy in women. Ideally, prevention or very early detection of breast cancer reduce both morbidity and mortality of the disease. Effective use of tumour markers might permit the application of screening and treatment more efficiently. MATERIALS AND METHODS: Immunohistochemical study was done in 39 cases of invasive breast carcinoma for further sub-classification for predicting prognosis and therapeutic approach. The cases were evaluated for expression of cytokeratin Ck 8/18, ER & Her-2/neu and 28 cases were subjected to cytokeratin Ck 5/6 including especially the cases negative for all the three markers. RESULTS: ER was positive in seven cases with Her-2/neu being positive in eight cases and cytokeratin 8/18 positive in 33 cases. Cytokeratin 5/6 was positive in seven cases with four cases showing Ck 5/6 and Ck8/18 positivity, three cases showing positivity for Ck 5/6 only and three cases were negative for all the four markers. CONCLUSION: Thus classifying them to be luminal A, luminal B, combined, basal and null type respectively. Luminal A tmours show the best prognosis while luminal B tumours , combined , basal and null type are associated with less favouable outcome. Thus it is important to categorise them according to the cell of origin for prognosis and treatment.

Cytokeratin 5/6 expression in benign and malignant breast lesions.

BACKGROUND: Cytokeratin s (CK) are used for the fingerprinting of carcinomas in general. In breast tissue, the luminal epithelial cells express CK 8/18, CK 7 and CK 19, while basal/myoepithelial cells express CK 5/6, CK 14 and CK 17. MATERIAL AND METHODS: Immunohistochemical staining for cytokeratin 5/6 was applied on cell block sections of 23 cases of benign and 25 cases of malignant breast lesions using avidin biotin peroxidase technique. The distribution and intensity of staining was recorded and graded semiquantitatively. RESULT: All benign lesions showed positive immunoreaction, with the staining index varying from 6-9, except lactating adenoma. The malignant lesions comprised three cases of ductal carcinoma in situ (DCIS) and 22 cases of infiltrating ductal carcinoma, not otherwise specified, IDC (NOS). None of the DCIS cases showed a positive immunoreaction. Among the IDC (NOS) lesions, six cases of grade III breast carcinoma exhibited a positive immunohistochemical reaction, the staining index of which varied from 2-6. The staining reaction in the malignant lesions was only cytoplasmic and the intensity was significantly less than that of benign lesions. CONCLUSION: CK 5/6 expression breast carcinoma implies a 'basal like' molecular phenotype and is associated with poor prognosis. This antibody is also used as a component of panels to differentiate benign and malignant breast lesions.

Direct laryngoscopic observations in progressive hoarseness of voice.

Hoarseness is a common disorder of voice which indicates abnormality at the level of glottis. We present our study of forty cases of progressive hoarseness of voice who underwent direct laryngoscopy during a period of January 2003 to January 2004 in our hospital. The observations were recorded und the data was evaluated.

Proton MR spectroscopy after acute central nervous system injury: outcome prediction in neonates, infants, and children.

PURPOSE: To evaluate the usefulness of proton magnetic resonance (MR) spectroscopy in predicting 6-12-month neurologic outcome in children after central nervous system injuries. MATERIALS AND METHODS: Localized single-voxel, 20-msec-echo-time MR spectra (including N-acetylaspartate [NAA], choline [Ch], creatine and phosphocreatine [Cr]) were obtained in the occipital gray matter in 82 patients and 24 control patients. Patient age groups were defined as neonates (< or = 1 month [n = 23]), infants (1-18 months [n = 31]), and children (> or = 18 months [n = 28]). Metabolite ratios and the presence of lactate were determined. Linear discriminant analysis-with admission clinical data, proton MR spectroscopy findings, and MR imaging score (three-point scale based on severity of structural neuroimaging changes)-was performed to help predict outcome in each patient. Findings were then compared with the actual 6-12-month outcome assigned by a pediatric neurologist. RESULTS: Outcome on the basis of proton MR spectroscopy findings combined with clinical data and MR imaging score was predicted correctly in 91% of neonates and in 100% of infants and children. Outcome on the basis of clinical data and MR imaging score alone was 83% in neonates, 84% in infants, and 93% in children. The presence of lactate was significantly higher in patients with poor outcome than in patients with good-moderate outcomes in all three age groups (neonates, 38% vs 5%; infants, 87% vs 5%; children, 64% vs 10% [chi 2 test, P < .02]). In children with poor outcomes, NAA/Cr ratios were significantly lower in infants (P = .006) and children (P < .001), and NAA/Ch ratios were significantly lower in infants (P = .001) and neonates (P = .05). CONCLUSION: Findings at proton MR spectroscopy helped predict long-term neurologic outcomes in children after central nervous system injury.