PERBANDINGAN PAPARAN ASAP ROKOK KONVENSIONAL DAN ROKOK HERBAL PADA MENCIT (Mus musculus) TERHADAP PERBANDINGAN GAMBARAN HISTOLOGI PARU

The purpose of this study was to determine the comparison of conventional cigarette smoke and herbal cigarettes against histological damage in the lungs of mice. The research method uses a completely randomized design with 21 mice aged 8-12 weeks and divided into 3 treatment groups. Group P0 was not given exposure to cigarette smoke, group P1 was exposed to conventional cigarette smoke tar content was 39 mg nicotine 2 mg, group P2 was exposed to herbal cigarette smoke tar content was 41.60 mg nicotine 1.64 mg. On the 15th day the mice were deuthanated for pulmonary extraction and haematoxyline eosin staining was made. Data analysis using one way ANOVA. The results of this study obtained scoring lung damage to edema lesions in P0, P1, P2 Sig 0.246 (p> 0.05). Inflammation of inflammatory cells at P0, P1, P2 Sig 0.000 (p <0.05). Destruction of alveolar septum at P0, P1, P2 Sig 0.000 (p <0.05). The conclusion of this study is the difference in lung damage in mice in the form of edema, infiltration of inflammatory cells, destruction of the alveolar septum between exposure to conventional cigarette smoke and herbal cigarettes. The purpose of this study was to determine the comparison of conventional cigarette smoke and herbal cigarettes against histological damage in the lungs of mice. The research method uses a completely randomized design with 21 mice aged 8-12 weeks and divided into 3 treatment groups. Group P0 was not given exposure to cigarette smoke, group P1 was exposed to conventional cigarette smoke tar content was 39 mg nicotine 2 mg, group P2 was exposed to herbal cigarette smoke tar content was 41.60 mg nicotine 1.64 mg. On the 15th day the mice were deuthanated for pulmonary extraction and haematoxyline eosin staining was made. Data analysis using one way ANOVA. The results of this study obtained scoring lung damage to edema lesions in P0, P1, P2 Sig 0.246 (p> 0.05). Inflammation of inflammatory cells at P0, P1, P2 Sig 0.000 (p <0.05). Destruction of alveolar septum at P0, P1, P2 Sig 0.000 (p <0.05). The conclusion of this study is the difference in lung damage in mice in the form of edema, infiltration of inflammatory cells, destruction of the alveolar septum between exposure to conventional cigarette smoke and herbal cigarettes.

CARD11 Mutation Induces Oligoclonal Expansion of T-Cells, and Accelerates ATL Development in Combination with HBZ

Adult T-cell leukemia/lymphoma (ATL) is an aggressive peripheral T-cell lymphoma that develops in about 5% of human T-cell leukemia/lymphoma virus 1 (HTLV-1) carriers. In addition to viral oncogenes, namely tax and HTLV-1 bZIP factor (HBZ), gene mutations, highly enriched for T-cell receptor (TCR)-NF-kB signaling, should be involved in the development of ATL. Among gene mutations, mutation in CARD11, a cytoplasmic scaffolding protein required for TCR-induced NF-kB activation, was detected in 24% of ATL patients. Here we generated a mouse model for conditional expression of a human ATL-derived CARD11(E626K) gain-of-function mutant, and demonstrated CARD11 activation induced oligoclonal expansion of T-cell and infiltration to many organs. We also showed that expression of HBZ accelerated mutant CARD11-induced lymphoproliferative diseases. We introduced a human Card11(E626K) into the mouse genome at the ROSA26 locus. After crossing with CD4-Cre Tg, CARD11(E626K)CD4-Cre mice was obtained. In CD4+ cells from CARD11(E626K)CD4-Cre, the amount of cleaved BCL-10 and NF-kBp65 increased compared with those in WT CD4+cells, confirming the activation of NF-kB. About half of CARD11(E626K)CD4-Cre mice died on or after 6 months after birth. At 6 months, leukocytosis was observed in CARD11(E626K)CD4-Cre, and accordingly the number of CD4+ cells cells was about 1.43 times greater than those in WT mice. The most affected organ in CARD11(E626K)CD4-Cre mice was lung. Alveolar septum was thickened by infiltrated cells at 6 months, and worsened subsequently. CD3+ T-cell accumulated around capillary blood vessels, and had high proliferation indices (&gt;50%), as assayed by Ki-67 staining. CARD11(E626K)CD4-Cre mice developed lymphadenopathy (4/8 mice (50%) at 6M and 4/6 mice (66.7%) at 12M). Normal lymph node (LN) architecture was barely preserved, and medullary sinus was expanded with CD3+ T-cell. Some of them were positive for FoxP3, and had moderate proliferation indices (25%-50%). Among CD4+ T-cell, the proportion of naive T-cell (Tn) decreased, and that of effector/memory T-cell (Tem) and regulatory T-cell (Treg) increased compared to WT LNs. The proportion of Treg in CD4+ T-cell from LN of 6M- and 12M-old CARD11(E626K)CD4-Cre mice was 25% and 40%, respectively, which values were much larger than the normal range as 10-15%. We next examined the clonality of CD4+ cells in spleen and swollen LNs from CARD11(E626K)CD4-Cre mice. The clonality of the TCR repertoires of 20 individual Vb gene famines from Vb1-20 was assessed by a PCR. The clonality assay using the TCR repertoires exhibited an oligoclonal pattern in 4 of 5 splenic CD4+ cells, and 1 of 2 LN CD4+ cells from CARD11(E626K)CD4-Cre mice. To assess the effect of HBZ constant expression on CARD11(E626K)CD4-Cre mice, we generated HBZ Tg, in which HBZ cDNA was expressed under the CD4 promoter. Similar to the previous report (by Satou et al.), our HBZ Tg showed increased number of Tem, destroyed architecture of lung such as thickened alveolar septum by infiltrated cells and decreased alveolar space by edema, and the lymphadenopathy after 12M (66.7%). We then crossed CARD11(E626K)CD4-Creand HBZ Tg, and obtained the compound mice. The compound mice caused more aggressive lymphoproliferative diseases compared with CARD11(E626K)CD4-Cre mice. Most of compound mice died within 8M. At 6M, architecture of lung, kidney, spleen, and LN was destroyed. In lung, alveolar space of lung was scarcely observed caused of T-cell invasion. Alveolar septum was thickened with infiltrated cells, and CD3+ cells accumulated around capillary blood vessel. Some of them were positive for FoxP3, and indicated moderate proliferation indices (25-50%). T-cell invasion was also observed in kidney. Lymphadenopathy was detected in 6 of 9 (66.7%) with completely destroyed architecture, increment of the proportion of Fox3+ cells, and moderate proliferation indices (25-50%). The clonality assay using the TCR repertoires exhibited an oligoclonal pattern in 4 of 4 splenic CD4+ cells, and 2 of 2 LNs CD4+ cells from compound mice. These results suggest that CARD11 mutant-induced NFkB activation and constant HBZ expression may have similar effects, such as T-cell infiltration into organs and LN adenopathy, and that the simultaneous occurrence of both may have additive effects. Disclosures Sugiyama: Chordia Therapeutics, Japan.: Current Employment. Morishita:Chordia Therapeutics Inc.: Current Employment, Current equity holder in private company. Shimoda:Perseus Proteomics: Research Funding; PharmaEssentia Japan: Research Funding; AbbVie Inc.: Research Funding; Astellas Pharma: Research Funding; Merck & Co.: Research Funding; CHUGAI PHARMACEUTICAL CO., LTD.: Research Funding; Kyowa Hakko Kirin Co., Ltd.: Research Funding; Pfizer Inc.: Research Funding; Otsuka Pharmaceutical: Research Funding; Asahi Kasei Medical: Research Funding; Japanese Society of Hematology: Research Funding; The Shinnihon Foundation of Advanced Medical Treatment Research: Research Funding; Celgene: Honoraria; Shire plc: Honoraria; Novartis: Honoraria, Research Funding; Takeda Pharmaceutical Company: Honoraria; Bristol-Myers Squibb: Honoraria.

Endothelial glycocalyx shedding in the acute respiratory distress syndrome after flu syndrome

Background Scientific evidence indicates that endothelial glycocalyx (EG) shedding contributes to the pathophysiological installation of acute respiratory distress syndrome (ARDS) after bacterial sepsis. The aim was to evaluate the EG shedding in ARDS installation after flu syndrome. Methods This cross-sectional study included patients with flu syndrome during the influenza outbreak divided into two groups: patients with and without ARDS. Healthy subjects without flu syndrome were included in a control group. We measured EG damage biomarkers (hyaluronan, syndecan-1) and endothelial cell injury biomarker (soluble thrombomodulin) during the first medical evaluation. Histological assessment of the perimeter of the hyaline membrane and the number of neutrophils infiltrated in the alveolar septum was performed in patients who died. Results ARDS group had 30 patients (44 ± 16 years old, 57% men), the non-ARDS group had 36 patients (39 ± 17 years old, 42% men), and the control group had 35 individuals (44 ± 9 years old, 51% men). Hyaluronan levels were significantly higher in the ARDS group than the two groups [31 ng/ml (interquartile range-IQR 12–56) vs. 5 ng/ml (IQR 3–10) vs. 5 ng/ml (IQR 2–8); p < 0.0001]. Hyaluronan levels above 19 ng/ml in patients with flu syndrome were associated with a significant increase in 28-day mortality rate: relative risk (RR): 6.95; (95% confidence interval 1.88–25.67); p = 0.0017. A positive correlation was observed between hyaline membrane perimeter and soluble thrombomodulin levels (r = 0.89; p = 0.05) as well as between the number of neutrophils in the alveolar septum and hyaluronan levels (r = 0.89; p = 0.05). Conclusions Evidence of EG shedding was found in ARDS established after flu syndrome.

An Unusual Presentation of Congenital Lobar Emphysema

Congenital lobar emphysema is an uncommon bronchopulmonary malformation characterized by lobar overinflation and accompanying alveolar septum damage that leads to compression atelectasis of the lung parenchyma and displacement of mediastinal structures, with the resultant ventilation-perfusion mismatch. We present a case of a 33-year-old lady with progressive exertional dyspnea. Chest radiograph findings lead to the suspicion of congenital lobar emphysema, which was then confirmed by a computed tomography (CT) scan. This condition is most commonly identified in newborns, with very few cases being reported in adults. Lobectomy remains the treatment of choice and in general has good outcome.

Abstract 58: Intratracheal Delivery of Autologous Bone Marrow-derived Cells Attenuate Muscularization and Inflammation in Lungs of Monocrotaline-induced Pulmonary Hypertension Model Rats.

Background: Vascular remodeling and inflammation contribute to the pathogenesis and/or development of pulmonary hypertension. Bone marrow-derived cells (BMCs) include mesenchymal stem cells and progenitor cells play critical role in tissue repair as well as anti-inflammatory effects. Aim of this study was to investigate the effect of intratracheal delivered BMCs on monocrotaline (MCT)-induced muscuralization and inflammation in rat lung. METHODS: BMCs were isolated and cultured for 3 weeks. Seven days following MCT (60 mg/kg) treatment, DiI labeled autologous BMCs were intratracheally delivered to male Sprague-Dawley rats. Twenty-eight days following the MCT treatment, rats had hemodynamic studies. Hearts and lungs were removed to evaluate right ventricular (RV) hypertrophy and lung pathologies. Results: MCT treated rats had increased RV systolic pressure (RVSP), RV weight as well as wall thickness of small pulmonary arteries and alveolar septum thickness than those of placebo-treated control rats (all p values <0.01). Intratracheal delivery of autologous BMCs (1x106) 7 days after MCT prevented the increase in RVSP at 4 weeks compared with MCT alone (45.1 ± 2.23 versus 35.0 ± 3.84 mmHg, respectively; p<0.05). Furthermore, the intratracheal delivery of BMCs significantly attenuated the pulmonary vessel wall thickness (54.4 % ± 2.25 versus 17.8 % ± 2.69, p<0.01), and alveolar septum thickening (155.1 % ± 1.56 versus 114.2 % ± 1.16, p<0.01) accompanied with decreased numbers of inflammatory cells in perivascular areas and expression of proinflammatory cytokines compared with MCT alone. Immunohistochemical studies showed monocyte-like shaped DiI positive cells distributed in alveolar areas in BMCs delivered lungs, and some of those DiI positive cells expressed CD 68 and alpha smooth muscle cell actin. Conclusion: Intratracheal delivery of autologous BMCs prevented MCT-induced pulmonary hypertension by attenuation of vascular muscularization and inflammation in rat lung.

Prostaglandin F synthase is localized to contractile interstitial cells in bovine lung.

It was recently found that certain cells in the alveolar septum of the bovine lung are enriched in prostaglandin F (PGF) synthase. In this study we used immunohistochemical techniques at both light and electron microscopic levels to further characterize the PGF synthase-positive cells. By double immunofluorescence staining of bovine lung cryostat sections, the alveolar septal cells labeled by anti-PGF synthase antibody were also intensely labeled for cytoplasmic actin but not for alpha-smooth muscle actin. This labeling pattern suggests that the PGF synthase-positive cells in the septum are "contractile interstitial cells," which resemble conventional fibroblasts but characteristically contain prominent bundles of actin filaments. Immunogold electron microscopy of ultra-thin frozen sections of bovine lung showed that alveolar interstitial cells extending long cytoplasmic processes and closely associated with alveolar capillaries were intensely labeled for PGF synthase. Capillary endothelial cells, alveolar epithelial cells, and some fibroblastic cells were devoid of labeling. On the basis of these findings, we conclude that PGF synthase is specifically expressed in contractile interstitial cells within the alveolar septum. The protein may be a useful marker for contractile interstitial cells, whose physiological function and role in various pathological conditions have not been characterized in detail.