Inflammatory Caspases Drive Pyroptosis in Acute Lung Injury

Acute lung injury (ALI), a critical respiratory disorder that causes diffuse alveolar injury leads to high mortality rates with no effective treatment. ALI is characterized by varying degrees of ventilation/perfusion mismatch, severe hypoxemia, and poor pulmonary compliance. The diffuse injury to cells is one of most important pathological characteristics of ALI. Pyroptosis is a form of programmed cell death distinguished from apoptosis induced by inflammatory caspases, which can release inflammatory cytokines to clear cells infected by pathogens and promote monocytes to reassemble at the site of injury. And pyroptosis not only promotes inflammation in certain cell types, but also regulates many downstream pathways to perform different functions. There is increasing evidence that pyroptosis and its related inflammatory caspases play an important role in the development of acute lung injury. The main modes of activation of pyroptosis is not consistent among different types of cells in lung tissue. Meanwhile, inhibition of inflammasome, the key to initiating pyroptosis is currently the main way to treat acute lung injury. The review summarizes the relationship among inflammatory caspases, pyroptosis and acute lung injury and provides general directions and strategies to conduct further research.

Korelasi Marshall CT score sebagai prediktor mortalitas pada penderita cedera kepala di RSUD dr Abdul Aziz Singkawang

Latar Belakang: Cedera kepala termasuk masalah global dan salah satu penyebab terbanyak kematian dan kecacatan diseluruh dunia. Insiden cedera kepala banyak terjadi pada usia produktif, yaitu pada usia 15-24 tahun. Di Indonesia kasus trauma merupakan penyebab kematian terbanyak ke-4 setelah stroke, tuberkulosis dan hipertensi. Insiden terjadinya cedera kepala di Indonesia relatif tinggi, terutama cedera kepala akibat kecelakaan lalu lintas yang mencapai angka 19,6%. Di Kalimantan Barat sendiri jumlah korban meninggal akibat kecelakaan lalu lintas terjadi sebanyak 560 orang pada tahun 2013, 550 orang pada tahun 2014 dan 470 orang pada tahun 2015. Sedangkan insiden kecelakaan lalu lintas di Kota Singkawang tercatat 211 kasus pada tahun 2012, 168 kasus pada tahun 2013, 120 kasus pada tahun 2014, 110 kasus pada tahun 2015 dan terdapat 71 kasus pada tahun 2016. Metode: Penelitian ini bersifat analitik dengan menggunakan pendekatan potong lintang. Dilakukan di RSUD Dr Abdul Aziz Kota Singkawang. Penelitian ini menggunakan total sampling dari seluruh rekam medis pasien bedah saraf dengan cedera kepala sedang dan berat diRSUD Abdul Aziz Kota Singkawang. Data yang diambil adalah hasil penilaian CT Scan pasien menggunakan Marshall CT Score dan penilaian hasil keluar, yang diambil berdasarkan rekam medis pasien. Sampel minimal yang diambil sebanyak 30 sampel yang sesuai dengan kriteria inklusi dan eksklusi. Hasil: terdapat korelasi positif yang sangat kuat antara Marshall CT Score dan mortalitas pada pasien cedera kepala sedang dan berat dengan nilai r mencapai 0.943 dari 66 data. Kesimpulan: Terdapat korelasi positif yang sangat kuat antara Marshall CT Score dan mortalitas, yang mana semakin tinggi nilai Marshall maka semakin tinggi angka mortalitas. Nilai GCS yang terbanyak pada subjek penelitian adalah 12. Nilai Marshall CT Score terbanyak yaitu kategori diffuse injury II, dan persentase mortalitas tertinggi pada kategori non evacuated mass lesion. Angka mortalitas pada pasien cedera kepala sedang dan berat adalah 26 orang.

Hubungan Kadar Interleukin 6 Serum dan Klasifikasi CT Marshal pada Penderita Cedera Otak Berat Akibat Trauma

Increase of interleukin 6 (IL-6) level occurs in the brain after traumatic brain injury (TBI), however, studies about IL-6 as a prodictor based on CT-scan is still limited. This study was aimed to evaluate the relationship between serum IL-6 level and CT Marshall classification in patients with severe TBI. This was an observational study with a cross sectional design. There were 20 patients with severe TBI admitted at the Emergency Surgery Installation of Prof. Dr. R. D. Kandou Hospital Manado in this study. CT-scan was performed on them to determine the CT Marshall classification and to categorize the hemorrhage location (extra-axial, intra-axial, both), hemisphere (midline/diffuse, dextral, sinistral), and area (frontal, parietal, temporal, occipital, multiple). Venous blood sample used for IL-6 examination was drawn less than 24 hours after trauma. The results showed that mean IL-6 level was 22.0060 pg/mL (SD 4.64494 pg/mL). Patients were distributed relatively uniform in 4 detected categories (diffuse injury II, III, V, and VI) of CT Marshall classification. Final regression model consisted of IL-6, age, and temporal injury as predictors. The Spearman coefficient correlation showed rs = -0.005 (P=0.491). Conclusion: There was no significant relationship between serum Il-6 level and CT Marshall classification, albeit, both of them increased consistantly following the severity of TBI and could be potential predictors to determine the prognosis of severe TBI patients.Keywords: IL-6, CT Marshall, severe TBIAbstrak: Pada cedera otak berat akibat trauma (COBT) terjadi peningkatan ekspresi IL-6 di otak namun penelitian mengenai kemampuannya untuk memrediksi hasil berdasarkan klasifikasi CT scan masih terbatas. Penelitian ini bertujuan untuk mengevaluasi hubungan antara kadar IL-6 serum dan klasifikasi CT Marshall pada pasien dengan COBT. Jenis penelitian ialah observasional dengan desain potong lintang. Hasil penelitian mendapatkan 20 pasien yang dirawat dengan COBT di IRDB RSUP Prof. Dr. R. D. Kandou Manado. CT-scan segera digunakan untuk menentukan klasifikasi CT Marshall dan untuk mengategorikan lokasi (ekstra-aksial, intra-aksial, keduanya), belahan (garis tengah/difus, dekstra, sisnitra), dan area otak yang terlibat dalam cedera. Sampel darah vena untuk IL-6 diambil kurang dari 24 jam setelah trauma. Hasil penelitian mendapatkan rerata kadar IL-6 22,0060 pg/mL (SD 4,64494 pg/mL). Pasien didistribusikan relatif seragam dalam empat kategori yang terdeteksi (difus cedera II, III, V, dan VI) dari klasifikasi CT Marshall. Model regresi akhir terdiri dari IL-6, usia, dan cedera pada area temporal sebagai prediktor. Korelasi antara kadar IL-6 serum dan klasifikasi CT Marshall dianalisis dengan koefisien korelasi Spearman dan mendapatkan rs = -0,005 (P=0,491). Simpulan: Walaupun secara statistik tidak terdapat hubungan bermakna antara kadar Il-6 serum dan CT Marshall namun keduanya secara konsisten meningkat mengikuti COBT dan dapat menjadi prediktor potensial untuk menentukan prognosis pada pasien dengan COBT.Kata kunci: IL-6, CT Marshall, COBT

Using selective lung injury to improve murine models of spatially heterogeneous lung diseases

Many lung diseases, such as acute respiratory distress syndrome (ARDS), display significant regional heterogeneity, with patches of severely injured tissue adjacent to apparently healthy tissue. Current mouse models that aim to mimic ARDS generally produce diffuse injuries that cannot reproducibly generate ARDS’s regional heterogeneity. This deficiency prevents the evaluation of how well therapeutic agents reach the most injured regions, and precludes many regenerative medicine studies, since it is not possible to know which apparently healing regions suffered severe injury initially. Finally, these diffuse injury models must be mild to allow for survival, as their diffuse nature does not allow for residual healthy lung to keep an animal alive long enough for many drug and regenerative medicine studies. To solve all of these deficiencies of current animal models, we have created a simple and reproducible technique to selectively induce lung injury in specific areas of the lung. Our technique, catheter-in-catheter selective lung injury (CICSLI), involves guiding an inner catheter to a particular area of the lung and delivering an injurious agent mixed with nanoparticles (fluorescently and/or radioactively labeled) that can be used to track the location and extent of where the initial injury was, days later. Further, we demonstrate that CICSLI can produce a more severe injury than diffuse models, yet has much higher survival since CICSLI intentionally leaves undamaged lung regions. Collectively, these attributes of CICSLI will allow better study of how drugs act within heterogeneous lung pathologies and how regeneration occurs in severely damaged lung tissue, thereby aiding the development of new therapies for ARDS and other lung diseases.

Coma

Coma is a pathological state of unconsciousness from which a patient cannot be roused to wakefulness by stimuli, and reflects dysfunction of the brainstem reticular system and its thalamic projections (the neuronal basis of wakefulness), or diffuse injury of both cerebral hemispheres. A unilateral lesion of a cerebral hemisphere (e.g. haemorrhagic stroke) will not cause coma unless there is secondary compression of the contralateral hemisphere or brainstem. Coma is a medical emergency, because a comatose patient is at high risk of permanent brain injury or death, caused either by the underlying disorder or the secondary effects of coma. Stabilization of the airway, breathing, and circulation, and exclusion of hypoglycaemia are the first priorities, before diagnosis is explored further. Clinical assessment together with neuroimaging will usually identify the likely cause or causes. The clinical approach to diagnosis and management of the comatose patient is described in this chapter.

Abstract 20: Caridomoyocyte Biology Revealed by Fluorescence Ubiquitination-based Cell-cycle Indicators

Existing myocyte contribution to new myocyte formation remains an active area of investigation. Novel experimental methodology is needed to faithfully label cardiomyocyte cell-cycle activity after birth and following injury. The Fluorescence Ubiquitination Cell Cycle Indicator (FUCCI) system can be used to aid visualization of cell cycle activity and progression by monitoring the inverse oscillation dynamics of fluorescently tagged cell cycle fusion proteins AzG-hGeminin and mKO2-hCdt1. Using this system, we hypothesize that cardiomyocytes retain the capacity to cycle throughout postnatal development and re-enter the cell cycle following acute myocardial infarction injury (MI). A novel cardiac specific FUCCI transgenic mouse model, αMHC-FUCCI, was developed to study cell-cycle dynamics of cardiomyocytes. αMHC-FUCCI hearts were collected throughout postnatal (PN) development to examine cardiomyocyte cell-cycle. Similarly, adult αMHC-FUCCI mice were subjected to MI, injected daily with BrdU and harvested at 3, 7, 10, 14 and 21 days post-MI for further analysis. Peak incidence of single mKO2-hCdt1 (7%, G 1 ) and AzG-hGem (2%, S/G 2 /M) fluorescence in cardiomyocytes occurs at PN7 and decreases over time as as confirmed by colocalization with BrdU and/or mitotic marker phospho-histone 3. Interestingly, continued mitotic activity exists at PN14 as observed by AzG+/pH3+ myocytes and concurrent mKO2+/AzG+ fluorescence is observed in 60% of adult myocardium by at one month. Together, these results indicate cardiac myocytes remain active at least two weeks after birth and transition into a G1/S phase as opposed to a mitotic exit (G0) as adults. Intriguingly, BrdU+ label is only detected in the non-myocyte interstitial population in and around the border zone through the first two weeks post-MI. BrdU+/AzG+ and or/mKO+ myocytes are detectable at 21days post-MI, indicating a lag in cardiomyocyte cell cycle re-entry. These results suggest myocytes retain the ability to re-enter the cell cycle at low levels three weeks post-MI. Future studies will analyze cardiomyocyte cell-cycle biology in response to diffuse injury and will further elucidate the mechanism behind myocardial regeneration.

Antenatal dexamethasone before asphyxia promotes cystic neural injury in preterm fetal sheep by inducing hyperglycemia

Antenatal glucocorticoid therapy significantly improves the short-term systemic outcomes of prematurely born infants, but there is limited information available on their impact on neurodevelopmental outcomes in at-risk preterm babies exposed to perinatal asphyxia. Preterm fetal sheep (0.7 of gestation) were exposed to a maternal injection of 12 mg dexamethasone or saline followed 4 h later by asphyxia induced by 25 min of complete umbilical cord occlusion. In a subsequent study, fetuses received titrated glucose infusions followed 4 h later by asphyxia to examine the hypothesis that hyperglycemia mediated the effects of dexamethasone. Post-mortems were performed 7 days after asphyxia for cerebral histology. Maternal dexamethasone before asphyxia was associated with severe, cystic brain injury compared to diffuse injury after saline injection, with increased numbers of seizures, worse recovery of brain activity, and increased arterial glucose levels before, during, and after asphyxia. Glucose infusions before asphyxia replicated these adverse outcomes, with a strong correlation between greater increases in glucose before asphyxia and greater neural injury. These findings strongly suggest that dexamethasone exposure and hyperglycemia can transform diffuse injury into cystic brain injury after asphyxia in preterm fetal sheep.