Respiratory Tree Research Articles

Long-term results of PRRT in advanced bronchopulmonary carcinoid.

Peptide receptor radionuclide therapy (PRRT) for the treatment of neuroendocrine tumours (NET) has been explored for almost two decades, but there are still few trials that have exclusively investigated well-differentiated and moderately differentiated NET arising from the respiratory tree. Thus, the aim of this study was to explore the outcome in patients affected by bronchopulmonary carcinoid (BPC) following PRRT. We retrospectively analysed 114 patients with advanced stage BPC consecutively treated with PRRT at the European Institute of Oncology, Milan, from 1997 to 2012 and followed until October 2014. The objective responses, overall survival (OS) and progression-free survival (PFS) were rated, and three different PRRT protocols ((90)Y-DOTATOC vs. (177)Lu-DOTATATE vs. (90)Y-DOTATOC + (177)Lu-DOTATATE) were compared with regard to their efficacy and tolerability. The median OS (evaluated in 94 of the 114 patients) was 58.8 months. The median PFS was 28.0 months. The (177)Lu-DOTATATE protocol resulted in the highest 5-year OS (61.4%). Morphological responses (partial responses + minor responses) were obtained in 26.5% of the cohort and were associated with longer OS and PFS. The (90)Y-DOTATOC + (177)Lu-DOTATATE protocol provided the highest response rate (38.1%). Adverse events were mild in the majority of patients. However, haematological toxicity negatively affected survival. No severe (grade 3/4) serum creatinine increase was observed. Patients treated with (90)Y-DOTATOC alone more frequently showed a mild/moderate decrease in renal function. In patients treated with chemotherapy before PRRT had a shorter OS and PFS, and a higher risk of developing nephrotoxicity. In a large cohort of patients with advanced BPC treated in a "real-world" scenario and followed up for a median of 45.1 months (range 2-191 months), PRRT proved to be promising in prolonging survival and delaying disease progression. Despite the potential selection biases, considering the risk-benefit ratio, (177)Lu-DOTATATE monotherapy seems the best option for PRRT. Our results indicate that the use of PRRT in earlier stages of the disease could provide a more favorable outcome.

The first echinoderm poly-U-binding factor 60 kDa (PUF60) from sea cucumber (Stichopus monotuberculatus): Molecular characterization, inducible expression and involvement of apoptosis.

Poly-U-binding factor 60 kDa (PUF60), also known as Ro RNA binding protein (RoBPI) and FBP interacting repressor (FIR), is a multifunctional protein that is involved in a variety of nuclear processes including pre-mRNA splicing, apoptosis and transcription regulation. In this study, the first echinoderm PUF60 named StmPUF60 was identified from sea cucumber (Stichopus monotuberculatus). The StmPUF60 cDNA is 4503 bp in length, containing a 5'-untranslated region (UTR) of 34 bp, a 3'-UTR of 2963 bp and an open reading frame (ORF) of 1506 bp that encoding a protein of 501 amino acids with a deduced molecular weight of 54.15 kDa and a predicted isoelectric point of 5.15. The putative StmPUF60 protein possesses all the main characteristics of known PUF60 proteins, including two RNA recognition motifs (RRM1 and RRM2), a C-terminal PUMP domain and two conserved nucleic acid-binding ribonucleoprotein sequences (RNP1 and RNP2). For the gene structure, StmPUF60 contains nine exons separated by eight introns. In addition, the highest level of StmPUF60 mRNA expression was noticed in the gonad, followed by coelomocytes, intestine, respiratory tree and body wall. In in vivo experiments, the expression of StmPUF60 mRNA in coelomocytes and intestine was significantly up-regulated by lipopolysaccharides (LPS) challenge, suggesting that the sea cucumber PUF60 might play critical roles in the innate immune defense against bacterial infections. Moreover, we further confirmed that overexpressed StmPUF60 could induce apoptosis, and this function of StmPUF60 may be one of the innate immune defense mechanisms for sea cucumber against pathogen infections.

The effects of dietary lead on growth, bioaccumulation and antioxidant capacity in sea cucumber, Apostichopus japonicus.

Three different diets amended with lead nitrate [Pb(NO3)2] (100, 500 and 1000mg Pb/kg dry weight) and a Pb-free control diet (1.03mg Pb/kg dry weight) were fed to sea cucumber (Apostichopus japonicus) for 30 days. The patterns of Pb accumulation over time were determined in various tissues (body wall, intestine and respiratory tree), as well as growth performance and antioxidant enzymes activities. Pb accumulation in body wall and intestine increased with time in all dietary Pb treatments. When fed the highest Pb diet, the body wall exhibited the greatest Pb burden (16.37mg Pb/kg tissue wet weight), while Pb content in the intestine (2.68mg Pb/kg tissue wet weight) and the respiratory tree (1.78mg Pb/kg tissue wet weight) were lower than Pb content in the body wall by day 30. The body weight gain (BWG), specific growth rate (SGR) and survival rate (SR) had not been affected by 30 days oral administration of Pb supplemented diet. However, the antioxidant enzymes activities [superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px)] of test groups were lower than control group in body wall and malondialdehyde (MDA) concentration in the body wall was opposite after 30 days in sea cucumbers. In summary, this work reports toxic effects in sea cucumber, A. japonicus, after dietary exposure to Pb.

Identification and Characterization of MicroRNAs from Longitudinal Muscle and Respiratory Tree in Sea Cucumber (Apostichopus japonicus) Using High-Throughput Sequencing.

MicroRNAs (miRNAs), as a family of non-coding small RNAs, play important roles in the post-transcriptional regulation of gene expression. Sea cucumber (Apostichopus japonicus) is an important economic species which is widely cultured in East Asia. The longitudinal muscle (LTM) and respiratory tree (RPT) are two important tissues in sea cucumber, playing important roles such as respiration and movement. In this study, we identified and characterized miRNAs in the LTM and RPT of sea cucumber (Apostichopus japonicus) using Illumina HiSeq 2000 platform. A total of 314 and 221 conserved miRNAs were identified in LTM and RPT, respectively. In addition, 27 and 34 novel miRNAs were identified in the LTM and RPT, respectively. A set of 58 miRNAs were identified to be differentially expressed between LTM and RPT. Among them, 9 miRNAs (miR-31a-3p, miR-738, miR-1692, let-7a, miR-72a, miR-100b-5p, miR-31b-5p, miR-429-3p, and miR-2008) in RPT and 7 miRNAs (miR-127, miR-340, miR-381, miR-3543, miR-434-5p, miR-136-3p, and miR-300-3p) in LTM were differentially expressed with foldchange value being greater than 10. A total of 14,207 and 12,174 target genes of these miRNAs were predicted, respectively. Functional analysis of these target genes of miRNAs were performed by GO analysis and pathway analysis. This result provided in this work will be useful for understanding biological characteristics of the LTM and RPT of sea cucumber and assisting molecular breeding of sea cucumber for aquaculture.

Three members in JAK/STAT signal pathway from the sea cucumber Apostichopus japonicus: Molecular cloning, characterization and function analysis.

The JAK/STAT signal transduction pathway plays a critical role in host defense against bacterial infections. In the present study, we firstly cloned the full-length cDNAs of three molecules in JAK/STAT cascade, STAT5, FOXP and SOCS2, from sea cucumber Apostichopus japonicus (denoted as AjSTAT5, AjFOXP, AjSOCS2, respectively) and investigated their immune functions towards Vibrio splendidus infection and LPS exposure. The AjSTAT5 cDNA was composed of 2643 bp consisting of 787 amino acid residues which included protein interaction domain, STAT-α domain, DNA binding domain and SH2 domain. The putative AjFOXP contained a ZnF_C2H2 domain, the leucine zipper-like domain and FH domain, all of which were thought to be the representative characteristics of FOXP subfamily. The deduced amino acids sequence of AjSOCS2 included an SH2 domain and SOCS box domain similar to vertebrate SOCS counterparts. Phylogenetic trees further supported that all these three identified proteins belonged to novel members of JAK/STAT signal pathway in sea cucumber. Tissue specific expression analysis showed that three genes were ubiquitously expressed in all examined tissues. AjSTAT5 and AjFOXP were both dominantly expressed in intestine, tentacle and respiratory tree, and weak in muscle. In contrary, the peak expression of AjSOCS2 was observed in muscle and lowest in respiratory tree. The V. splendidus challenge and LPS exposure could both significantly up-regulate the mRNA expression of three genes, in which AjSOCS2 showed opposite expression trends to those of AjSTAT5 and AjFOXP. Silencing the AjSTAT5 by siRNA depressed the AjFOXP expression, but induced the expression level of AjSOCS2, revealing that AjSTAT5 might directly modulate AjFOXP, and AjSOCS2 function primarily by acting as a potent inhibitor involve in JAK/STAT pathway. The present study would expand our understanding on JAK/STAT signaling transduction pathway in modulating the innate immune responses of sea cucumber.

Identification and characterization a novel transcription factor activator protein-1 in the sea cucumber Apostichopus japonicus.

The transcription factor activator protein-1 (AP-1) is an important gene expression regulator with typical Jun and region-leucine zipper (bZIP) domains and can respond to a plethora of physiological and pathological stimulus. In this study, we identified a novel AP-1 gene in Apostichopus japonicus by transcriptome sequencing and RACE approaches (designated as AjAP-1). The full-length of AjAP-1 was of 2944 bp including a 5' untranslated region (UTR) of 201 bp, a 3' UTR of 1753 bp and a putative open reading frame of 990 bp encoding a polypeptide of 329 amino acid residues. Two representative domains of Jun and bZIP as well as two nuclear localization signals (NLSs) were also detected in deduced amino acid of AjAP-1. Spatial distribution expression indicated that AjAP-1 was ubiquitously expressed in all examined tissues with predominant expression in the body wall, moderate in the tube feet, respiratory tree and colemocytes and slightly weak in the intestine and longitudinal muscle. Time-course expression analysis in intestine and coelomocytes revealed that AjAP-1 both reached its peak expression at 4 h after Vibrio splendidus challenge with a 2.6 and 8.2-fold increase compared to their control groups, respectively. Taken together, all these results suggested that AjAP-1 was a novel immune factor and might be involved in the processes of anti-bacteria response in sea cucumber.

Molecular characterization of two novel molecular chaperones in bacterial-challenged Apostichopus japonicus

Molecular chaperones of 78kDa glucose-regulated protein (GRP78) and protein disulfide isomerase (PDI) are involved in protein folding and assembly in the endoplasmic reticulum (ER). Increasing evidences also suggest that these two molecules play an important role in immune response. In the present study, we cloned and characterized GRP78 and PDI genes from Apostichopus japonicus by RNA-seq and RACE approaches (designated as AjGRP78 and AjPDI, respectively). The AjGRP78 cDNA was of 2355bp including an open reading frame (ORF) of 2013bp encoding a protein of 670 amino acids with three heat shock protein 70 (HSP70) family signatures. AjGRP78 contained a 23-amino acid signal peptide at the N-terminus and a HDEL motif at the C-terminus, which supported the location of the protein in the ER. The full length cDNA of AjPDI was of 1893bp with a 5′ untranslated region (UTR) of 153bp, a 3′ UTR of 228bp and an ORF of 1512bp encoding a protein of 503 amino acids. A 17-amino acid signal peptide, two thioredoxin domains with two active sites of CGHC, and KDEL retention signal were totally conserved in the deduced amino acid of AjPDI. Phylogenic analysis and multiple alignments have shown that both genes shared remarkably higher degree of structural conservation and sequence identities with other counterparts from invertebrates and vertebrates, further supporting that the two proteins were novel members of molecular chaperone family. Spatial expression analysis revealed that AjGRP78 mRNA transcripts were dominantly expressed in the tentacle, while AjPDI mRNA levels were abundant in the muscle, intestine and respiratory trees. For Vibrio splendidus challenged sea cucumber, the peak expression of AjGRP78 and AjPDI mRNAs in coelomocytes were detected at 24h with 1.73-fold increase and at 6h with 1.83-fold increase compared with the control group, respectively. Similarly, a significant increase in the relative mRNA levels of AjGRP78 and AjPDI was also identified in 1μgmL−1 LPS exposed primary cultured coelomocytes. These results collectively suggested that AjGRP78 and AjPDI were ER chaperones of A. japonicus, of which expression is induced upon bacterial infection.

Critical roles of sea cucumber C-type lectin in non-self recognition and bacterial clearance.

C-type lectin is one important pattern recognition receptor (PRR) that plays crucial roles in multiple immune responses. A C-type lectin from sea cucumber Apostichopus japonicus (AjCTL-1) was characterized in the present study. The amino acid sequence of AjCTL-1 shared high similarities with other C-type lectins from invertebrates and vertebrates. The C-type lectin domain (CTLD) of AjCTL-1 contained a Ca(2+)-binding site 2 and four conserved cysteine residues. AjCTL-1 mRNA expression patterns in tissues and after bacterial challenge were then analysed. Quantitative PCR revealed that AjCTL-1 mRNA was widely expressed in the tested tissues of healthy sea cucumber. The highest expression level occurred in gonad followed by body wall, coelomocytes, tentacle, intestinum and longitudinal muscle, and the lowest expression level was in respiratory tree. AjCTL-1 mRNA expression in coelomocytes was significantly induced by gram-negative Listonella anguillarum and gram-positive Micrococcus luteus, with different up-regulation patterns post-challenge. Recombinant AjCTL-1 exhibited the ability to bind peptidoglycan directly, agglutinate M. luteus, Staphylococcus aureus and Escherichia coli, in a Ca(2+)-dependant manner, and enhance the phagocytosis of coelomocytes against E. coli in vitro. The results indicated that AjCTL-1 could act as a PRR in Apostichopus japonicus and had critical roles in non-self recognition and bacterial clearance against invading microbes.

The uptake of dissolved organic material by the sea cucumber Parastichopus californicus (Stimpson) and its potential role in visceral regeneration

The sea cucumber Parastichopus californicus atrophies and regenerates its internal organs on an annual cycle. Previous research has suggested that during the period of maximal regeneration (January–March) no movement or feeding occurs; however metabolic rate is doubled compared to non-regenerative periods (June–August). Uptake of dissolved organic material (DOM) from the aqueous environment could provide an external source of biosynthetic materials for regeneration of the internal organs. P. californicus were collected in the summer (actively moving and feeding, not degenerating or regenerating), winter (inactive, degenerating), and spring (inactive, regenerating). Animals were incubated in seawater containing a mixture of amino acids labeled with 15N and 13C. Incorporation of 15N (summer, winter, spring) and 13C (summer, spring) by the respiratory trees was significantly higher than incorporation by other types of tissues. Additionally, dorsal and ventral epidermis, oral tentacles, digestive tract, muscle, and tube feet incorporated 15N and/or 13C during at least one season. These results demonstrate that certain tissues, especially the respiratory trees, of P. californicus assimilate free amino acids from the sea water which could be used as biosynthetic materials for regeneration. These results also indicate that the respiratory trees may have a role in nutrition as well as respiration.

Insights from unusual aspects of the 1918 influenza pandemic.

The 1918 influenza pandemic was the most lethal single event in modern history. Besides its mortality the 1918 pandemic was unusual for several reasons. It preferentially killed young adults from 20 to 40 y with a peak mortality at age 28 y. Mortality was highly variable with death rates varying by at least 10 fold within similar groups of citizens, soldiers, cities and islands. Secondary bacterial pneumonia following influenza was the overwhelming cause of death and not viral pneumonitis or acute lung injury. Clinical expressions of the 1918 pandemic were unusual with bleeding into the respiratory tree including epistaxis and dark blue cyanotic skin. The 1918 influenza virus apparently ceased circulation in the human population in the early 1920s but continued to evolve in pigs. Immunizations using viruses from 1918 and 2009 can cross-protect laboratory animals even though the human mortality outcomes were very different between the first pandemics of the 20th and 21st centuries. Unusual aspects of historical epidemics may help to reconstruct what actually occurred in 1918 and thus better prepare for the next pandemic.

Posterior regeneration following fission in the holothurian Cladolabes schmeltzii (Dendrochirotida: Holothuroidea).

The regeneration of the posterior portion of the body after fission was studied in the holothurian Cladolabes schmeltzii using electron microscopy methods. Following fission, the aquapharyngeal complex, gonad and anterior portion of the first descending part of the intestine remain in the anterior fragment of the body. The entire regeneration process is divided into five stages. In the first three stages, the digestive system and damaged ends of the longitudinal muscle bands regenerate. The intestine is formed through the rearrangement and growth of the remaining portion of the first descending part of the intestine. The gut anlage grows down the mesentery and joins the regenerating cloaca. The cloaca is formed from two sources: its posterior portion appears as a result of immersion of the epidermis, while the anterior portion develops from the terminal segment of the growing intestine. Regeneration of muscles progresses in the typical manner for echinoderms: through immersion and myogenic transformation of the coelomic epithelium. Respiratory trees appear in animals when the growth of the external part of the body has begun (fourth stage). They are formed as an outgrowth of the dorsal wall of the anterior portion of the cloaca. It was concluded that regeneration of the posterior portion of the body in the holothurian C. schmeltzii following fission is realized through morphallactic rearrangements of the remaining parts of organs. The main mechanism through which the digestive, respiratory, and contractile systems are formed is epithelial morphogenesis.

Identification, expression pattern and potential role of variable lymphocyte receptor Aj-VLRA from Apostichopus japonicus in response to bacterial challenge

The variable lymphocyte receptors (VLRs) are found in jawless vertebrates (agnathans), and specifically recognize bacteria and viruses via their leucine-rich repeats (LRRs). VLRs are believed to be adaptive immune response molecules. Echinoderms do not have adaptive immune systems; however, in the present study, a VLR cDNA named Aj-VLRA was cloned and characterized from sea cucumber, Apostichopus japonicus. The complete cDNA of Aj-VLRA was 3072 bp, including a 1995 bp open reading frame encoding 664 amino acids comprising LRR domains, a predicted transmembrane helix and an N-terminal signal peptide. As determined by quantitative real-time reverse transcription polymerase chain reaction, Aj-VLRA transcripts are ubiquitously expressed in the body wall, longitudinal muscles, intestine and respiratory tree of A. japonicus. The expression level of Aj-VLRA was upregulated after challenge with four common marine bacteria. In situ hybridization showed that the expression of Aj-VLRA was widely distributed in the four tissues, particularly in the cytoplasm of epidermal cells. Recombinantly expressed Aj-VLRA (including the LRR domains) could bind to bacteria including Micrococcus lysodeikticus (Gram+) and Vibrio anguillarum (Gram−). Collectively, the results suggested that Aj-VLRA is related to an innate immune response of A. japonicus.

Identification and expression patterns of extracellular matrix-associated genes fibropellin-ia and tenascin involved in regeneration of sea cucumber Apostichopus japonicus

Sea cucumbers have a strong regenerative capacity. Many important genes involved in the molecular mechanism of regeneration and associated with intercellular signaling pathways of regeneration have been identified. The product of the fibropellin-ia gene forms a layer known as the apical lamina that surrounds the sea cucumber embryo throughout development. Meanwhile, the tenascin gene displays highly restricted and dynamic patterns of expression in the embryo and is expressed in the adult during normal processes such as wound healing, nerve regeneration and tissue involution. In this study, we cloned for the first time full-length cDNAs of fibropellin-ia (1390bp, encoding a 199 amino acid protein) and tenascin (1366bp, encoding a 179 amino acid protein) from Apostichopus japonicus (designated Aj-fnia and Aj-tenascin, respectively) using rapid amplification of cDNA ends. The structures and characteristics of these two genes were analyzed bioinformatically, and their expression patterns associated with extracellular matrix remodeling in regeneration of A. japonicus were investigated by real-time PCR and in situ hybridization (ISH). Expression levels of Aj-fnia and Aj-tenascin in the regeneration tissues were higher than those in normal tissues. The highest expression levels of Aj-fnia and Aj-tenascin were shown in the intestine and respiratory tree on the 15th and 20th days after sea cucumbers were eviscerated. In the body wall, the highest expression levels of Aj-fnia and Aj-tenascin occurred at 35 and 45min during early regeneration and then emerged between 5 and 7days again during late regeneration after the body wall was injured. ISH analysis revealed expression of these genes in the body wall, longitudinal muscle, intestine and respiratory tree. These findings suggest that Aj-fnia and Aj-tenascin are crucial genes that play important roles in the regeneration of the sea cucumber.

Identification and characterization of a novel Foxo transcription factors in Apostichopus japonicus

The forkhead box O (Foxo) transcription factors are involved in multiple signaling pathways and play key roles in immunoregulation in vertebrates. In the present study, we firstly identified a novel Foxo gene in Apostichopus japonicus coelomocytes using transcriptome sequencing and RACE approaches (denoted as AjFoxo). The full-length cDNA of AjFoxo was of 2248 bp with a 5′ untranslated region (UTR) of 177 bp, a 3′ UTR of 367 bp and an ORF of 1704 bp encoding a polypeptide of 567 amino acid residues. The highly conserved forkhead domain was also identified in AjFoxo with remarkably higher degree of structural conservation. AjFoxo transcripts could be detected in all examined tissues with predominant expression in the coelomocytes and muscle, and slightly weak in the tissues of tentacle, intestine and respiratory trees. Concerning the time-course expression of AjFoxo in coelomocytes, the relative expression of AjFoxo was dramatically decreased to 0.44-fold at 48 h compared with that in the control group after Vibrio splendidus challenge, which was consistent with that of AjIκB. RNA interference of AjFoxo in primary coelomocytes also significantly depressed the relative expression of AjIκB with a 0.37-fold decrease compared with control group. Taken together, these results indicated that AjFoxo was a novel immune regulator and might be involved in the processes of anti-bacteria response in sea cucumber through activating the transcription of AjIκB.

Delayed presentation of post tuberculous broncho oesophageal fistula in an adult: Perioperative anaesthetic management.

Sir, A 42-year-old male had a recurrent cough with expectoration for last 6 months more with supine position and aggravated by the intake of food. He was referred for a pre-anaesthetic checkup for left pneumonectomy with repair of broncho oesophageal fistula (BEF). He had a history of parietal lobe abscess, drained twice with residual left-sided hemiplegia and focal seizures and had completed two courses of antituberculous treatment (ATT) 15 years back. The patient could lie down only in left lateral position. His investigations were unremarkable. Pulmonary function testing showed moderate impairment of lung function with moderately reduced vital capacity. X-ray and computerised tomography of the chest showed cystic bronchiectasis in left lower lobe. There were fibrosis and pleural thickening in bilateral lung fields with dilated oesophagus. Communicating tract between the lateral wall of the lower oesophagus and left lower lobe bronchus was seen at D-8 [Figure 1]. Post-inflammatory BEF was suggested in view of surrounding inflammatory changes and mediastinal lymphadenopathy. Barium swallow confirmed the same. Magnetic resonance imaging of the brain revealed post-operative changes with residual ring configuration in right posterior frontal lobe with moderate perilesional oedema. Figure 1 Computed tomography scan chest showing cystic bronchiectasis in left lower lobe and fistulous communication between oesophagus and left lower lobe bronchus The patient was accepted under American Society of Anaesthesiologists grade III. Antibiotics, chest physiotherapy, incentive spirometry, deep breathing exercises and all antiepileptic medications were continued till the morning of surgery. The patient was already on continuous Ryle's tube aspiration. After pre-medication, the thoracic epidural catheter was placed at T6-T7 interspace. Epidural test dose was followed by 3 mg of morphine. Monitoring included continuous electrocardiography, invasive blood pressure, end tidal carbon dioxide, SpO2, temperature, urine output and blood loss. After nasogastric tube aspiration and pre-oxygenation in the left lateral position, injection fentanyl 50 μg and sleep doses of thiopentone were given. As the patient went asleep, he was turned supine, and the cricoid pressure was applied. Injection rocuronium 50 mg was given to felicitate insertion of right sided double lumen tube (DLT) 41F. Intermittent positive pressure ventilation was avoided. Placement of DLT was confirmed clinically and fibreoptically. The left lung was isolated and collapsed immediately to prevent gastric distension and loss of lung volume. The patient was placed in right lateral position. Anaesthesia was maintained with nitrous oxide, oxygen, isoflurane, and rocuronium. Left pneumonectomy with excision of BEF track was done with blood loss of 2.5 L which was adequately replaced. A nasogastric tube was also placed. At closure, 6 ml of 0.125% bupivacaine was given through the epidural catheter. After extubation of trachea, the patient was shifted to intensive care unit for observation and made an uneventful recovery. BEF is rarely reported in the literature.[1] It may be congenital or acquired. Acquired causes include malignancy, trauma, oesophageal diverticulosis, infections such as syphilis, tuberculosis (TB) and actinomycosis. TB is the most common cause of infective BEF. The development of BEF in TB is related to mediastinal lymph node involvement.[2] Infection and subsequent adhesions between tracheobronchial lymph nodes and oesophagus with subsequent development of a traction diverticulum communicating with the respiratory tree are the mechanism for BEF formation. The aim during induction of anaesthesia is to avoid gas flow through the fistula which can cause gastric distension and impaired ventilation.[3] Positive pressure ventilation should be avoided. Spontaneous ventilation should be maintained during induction until gentle ventilation by mask provides an effective gas exchange. Double lumen intubation is required which protects the contralateral lung from contamination and provides the ability to ventilate it without applying positive pressure to the fistula.[3] Keeping the same in mind, rapid sequence induction was planned as insertion of DLT under spontaneous ventilation is difficult. Once placed, the diseased lung was isolated and kept collapsed. A surgical field fire is a known hazard during the surgery for BEF repair.[4] Avoiding the use of electrocautery while the fistula is open, using wet sponges, and decreasing the oxygen concentration in the gases before the division of fistula could reduce this incidence.[4] We instituted one lung ventilation with little loss of gases and a quick repair of the fistula to avoid the same. Post-tuberculous BEF is a rare entity and scarcely reported in the literature. BEF as a complication of TB can present at varied time intervals. There are reports of both immediate as well as a presentation in the chronic healing phase of TB.[1,2] Our patient received two complete courses of ATT 15 years back. BEF developed a decade later during the chronic healing phase of TB. Chronic suppuration not only damaged the left lung but also lead to the development of BEF. To conclude, prompt recognition and meticulous perioperative management lead to the successful outcome of our patient.

Difficult airway after late postoperative bleeding in a case of total thyroidectomy, tracheal resection and reconstruction.

Difficult airway after late postoperative bleeding in a case of total thyroidectomy, tracheal resection and reconstruction.

Expression responses of five cold tolerant related genes to two temperature dropping treatments in sea cucumber Apostichopus japonicus

Environmental conditions, including ambient temperature, play important roles in survival, growth development, and reproduction of the Japanese sea cucumber, Apostichopus japonicus. Low temperatures result in slowed growth and skin ulceration disease. In a previous study, we investigated the effect of low temperature on gene expression profiles in A. japonicus by suppression subtractive hybridization (SSH). Genes encoding Ferritin, Lysozyme, Hsp70, gp96, and AjToll were selected from a subtracted cDNA library of A. japonicus under acute cold stress. The transcriptional expression profiles of these genes were investigated in different tissues (coelomocyte, respiratory tree, intestine, longitudinal muscle) after exposure to acute and mild temperature dropping treatments. The results show that (1) the five cold-tolerance-related genes were found in all four tissues and the highest mRNA levels were observed in coelomocyte and respiratory tree; (2) under the temperature dropping treatments, three types of transcriptional regulation patterns were observed: primary suppression followed by up-regulation at −2°C, suppressed expression throughout the two treatments, and more rarely an initial stimulation followed by suppression; and (3) gene expression suppression was more severe under acute temperature dropping than under mild temperature dropping treatment. The five cold-tolerance-related genes that were distributed mainly in coelomocyte and respiratory tissues were generally down-regulated by low temperature stress but an inverse up-regulation event was found at the extreme temperature (−2°C).

DNA methylation levels analysis in four tissues of sea cucumber Apostichopus japonicus based on fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) during aestivation.

DNA methylation plays an important role in regulating transcriptional change in response to environmental stimuli. In the present study, DNA methylation levels of tissues of the sea cucumber Apostichopus japonicus were analyzed by the fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) technique over three stages of the aestivation cycle. Overall, a total of 26,963 fragments were amplified including 9112 methylated fragments among four sea cucumber tissues using 18 pairs of selective primers. Results indicated an average DNA methylation level of 33.79% for A. japonicus. The incidence of DNA methylation was different across tissue types in the non-aestivation stage: intestine (30.16%), respiratory tree (27.61%), muscle (27.94%) and body wall (56.25%). Our results show that hypermethylation accompanied deep-aestivation in A. japonicus, which suggests that DNA methylation may have an important role in regulating global transcriptional suppression during aestivation. Further analysis indicated that the main DNA modification sites were focused on intestine and respiratory tree tissues and that full-methylation but not hemi-methylation levels exhibited significant increases in the deep-aestivation stage.

Impact of extrafine formulations of inhaled corticosteroids/long-acting beta-2 agonist combinations on patient-related outcomes in asthma and COPD.

Asthma and chronic obstructive pulmonary disease (COPD) are among the most common chronic diseases worldwide, characterized by a condition of variable degree of airway obstruction and chronic airway inflammation. A large body of evidence has demonstrated the importance of small airways as a pharmacological target in these clinical conditions. Despite a deeper understanding of the pathophysiological mechanisms, the epidemiological observations show that a significant proportion of asthmatic and COPD patients have a suboptimal (or lack of) control of their diseases. Different factors could influence the effectiveness of inhaled treatment in chronic respiratory diseases: patient-related (eg, aging); disease-related (eg, comorbid conditions); and drug-related/formulation-related factors. The presence of multiple illnesses is common in the elderly patient as a result of two processes: the association between age and incidence of degenerative diseases; and the development over time of complications of the existing diseases. In addition, specific comorbidities may contribute to impair the ability to use inhalers, such as devices for efficient drug delivery in the respiratory system. The inability to reach and treat the peripheral airways may contribute to the lack of efficacy of inhaled treatments. The recent development of inhaled extrafine formulations allows a more uniform distribution of the inhaled treatment throughout the respiratory tree to include the peripheral airways. The beclomethasone/formoterol extrafine formulation is available for the treatment of asthma and COPD. Different biomarkers of peripheral airways are improved by beclomethasone/formoterol extrafine treatment in comparison with equivalent nonextrafine inhaled corticosteroids/long-acting beta-2 agonist (ICS/LABA) combinations. These improvements are associated with improved lung function and clinical outcomes, along with reduced systemic exposure to inhaled corticosteroids. The increased knowledge in the pathophysiology of the peripheral airways may lead to identify specific phenotypes of obstructive lung diseases that would mostly benefit from the treatments specifically targeting the peripheral airways.

Aerosol deposition doses in the human respiratory tree of electronic cigarette smokers

Aerosols from eight e-cigarettes at different nicotine levels and flavoring were characterized as particle number size distributions in the range 5.6–560 nm by FMPS and CPC. Results were used to provide dosimetry estimates applying the MMPD model.Particle number concentrations varied between 3.26 × 109 and 4.09 × 109 part cm−3 for e-liquids without nicotine and between 5.08 × 109 and 5.29 × 109 part cm−3 for e-liquids with nicotine. No flavor effects were detected on particle concentration data. Particle size distributions were unimodal with modes between 107–165 nm and 165–255 nm, for number and volume metrics, respectively.Averagely, 6.25 × 1010 particles were deposited in respiratory tree after a single puff. Highest deposition densities and mean layer thickness of e-cigarette liquid on the lung epithelium were estimated at lobar bronchi.Our study shows that e-cigarette aerosol is source of high particle dose in respiratory system, from 23% to 35% of the daily dose of a no-smoking individual.