GDF15 alleviates the progression of benign tracheobronchial stenosis by inhibiting epithelial-mesenchymal transition and inactivating fibroblasts.

Benign tracheobronchial stenosis (BTS) is a fatal and incurable disease. Epithelial repair and matrix reconstruction play an important role in the wound repair process. If the interstitial context is not restored and stabilized in time, it can lead to pathological fibrosis. Here we attempted to identify cytokines that are involved in promoting wound repair. Growth differentiation factor 15 (GDF15) is a cytokine secreted by tracheal epithelial cells, which is indispensable for the growth of epithelial cells and inhibits the overgrowth of fibroblasts. GDF15 can counteract transforming growth factor-ß (TGFß1) stimulation of epithelial-mesenchymal transition (EMT) in tracheal epithelial cells and inhibit fibroblast activation via the TGFß1-SMAD2/3 pathway. In a rat model of tracheal stenosis, GDF15 supplementation alleviated the degree of tracheal stenosis. These results suggest that GDF15 prevents fibroblast hyperactivation and promotes epithelial repair in injured trachea. GDF15 may be a potential therapy to improve benign tracheobronchial stenosis.

Molecular Mechanisms and Physiological Changes behind Benign Tracheal and Subglottic Stenosis in Adults.

Laryngotracheal stenosis (LTS) is a complex and heterogeneous disease whose pathogenesis remains unclear. LTS is considered to be the result of aberrant wound-healing process that leads to fibrotic scarring, originating from different aetiology. Although iatrogenic aetiology is the main cause of subglottic or tracheal stenosis, also autoimmune and infectious diseases may be involved in causing LTS. Furthermore, fibrotic obstruction in the anatomic region under the glottis can also be diagnosed without apparent aetiology after a comprehensive workup; in this case, the pathological process is called idiopathic subglottic stenosis (iSGS). So far, the laryngotracheal scar resulting from airway injury due to different diseases was considered as inert tissue requiring surgical removal to restore airway patency. However, this assumption has recently been revised by regarding the tracheal scarring process as a fibroinflammatory event due to immunological alteration, similar to other fibrotic diseases. Recent acquisitions suggest that different factors, such as growth factors, cytokines, altered fibroblast function and genetic susceptibility, can all interact in a complex way leading to aberrant and fibrotic wound healing after an insult that acts as a trigger. However, also physiological derangement due to LTS could play a role in promoting dysregulated response to laryngo-tracheal mucosal injury, through biomechanical stress and mechanotransduction activation. The aim of this narrative review is to present the state-of-the-art knowledge regarding molecular mechanisms, as well as mechanical and physio-pathological features behind LTS.

Histological features of complete tracheal rings in congenital tracheal stenosis.

PURPOSE: Congenital tracheal stenosis is a disease in which complete tracheal cartilage rings (CTCR) cause airway narrowing. Although tracheal cartilage malformation has been suggested as a cause of CTCR, no histological studies have been performed. Here, we report a comparison of the tissues from CTCR and normal tracheal cartilage. METHODS: Thirty-one infants who underwent slide tracheoplasty at our institution from May 2016 to August 2019 were included. Tissues from ten autopsy cases without tracheal lesions were used as controls. The survey items were tracheal cartilage cell density, cartilage thickness, and chondrocyte findings. RESULTS: The median cartilage cell density from cases was 23/125 × 125 µm2 and from controls was 23.5/125 × 125 µm2 (p = 0.90). The median cartilage thickness from cases was 689 µm and from controls was 840 µm (p = 0.11). Comparing the ventral and dorsal sides of the CTCR tissues, the cell density was significantly different (median ventral 23/125 × 125 µm2; median dorsal 19.5/125 × 125 µm2; p = 0.034). There were no significant findings in the chondrocytes of the CTCR tissues. CONCLUSION: CTCR tissues did not differ in cartilage density and thickness from normal tracheal cartilage.

LACHT syndrome (Mardini-Nyhan association) with tracheal stenosis in a Thai newborn.

LACHT syndrome, or Mardini-Nyhan association, is an ultra-rare disorder, diagnosed solely by the clinical characteristics of lung agenesis, complex cardiac defects, and thumb anomalies. Only 12 patients have been reported worldwide, and here, we report a new clinical diagnosis of LACHT syndrome. Our patient was a male full-term newborn with left lung agenesis, congenital heart defects including ventricular septal defect, right-sided aortic arch, with aberrant left subclavian artery and Kommerell diverticulum, as well as left preaxial polydactyly and hemivertebra. Our patient appears to be the second LACHT syndrome case to also suffer from tracheal stenosis, which has only been reported once before in conjunction with this syndrome. In light of this, tracheal stenosis may be a phenotype for LACHT syndrome.

[Etiology, diagnosis and treatment of cicatricial tracheal stenosis]. / Etiologiya, diagnostika i lechenie rubtsovogo stenoza trakhei.

Development of tracheal surgery was associated with introduction of fundamentally new procedures: two-level reconstruction, redo tracheal resection, tracheal resection with simultaneous dissection of tracheoesophageal fistula. There are combined and staged techniques when tracheal repair or endoscopic interventions are performed as a stage before circular resection of trachea. However, a single algorithm for prevention and correction of postoperative complications is still absent in tracheal surgery. Further development of tracheal surgery directly depends on introduction of preventive measures and analysis of adverse factors associated with increased risk of complications. In this regard, ongoing researches in this area are very perspective.

Death due to external compression of the trachea in a patient with multinodular hemorrhagic goiter.

In this paper we describe the case of an 81-year-old Caucasian female (142 cm tall, weighing 45 kg) who suffered from a multinodular goiter for approximately 40 years. Following the onset of a clinical condition characterized by acute respiratory failure, she was transported to the emergency room by ambulance, where she died within a few hours after admission. A recent cardiac examination showed the absence of risk factors for cardiovascular disease, sinus tachycardia with a heart rate of 131 bpm, negative objectivity for signs of cardiocirculatory failure, a blood pressure of 120/80 mmHg and modest exertional dyspnea. A recent hemochemical laboratory analysis showed a TSH value of 0.01microUI/mL, FT3 value of 4.76 pg/mL and FT4 value of 2.33 ng/mL, pointing to a pattern of hyperthyroidism, attributable to Basedow's goiter. Autopsy showed some peculiarities, and we came across two extremely rare findings; the thyroid gland had reached a very large size in relation to the patient's body mass (1510 g, in a patient of 142 cm and 45 kg), and the death of the patient was due to the development of a massive intra-thyroid hemorrhage that had caused acute external compression of the trachea. To the best of our knowledge this very rare event has not previously been reported in the international scientific literature.

Rabbit model of tracheal stenosis using cylindrical diffuser.

BACKGROUND AND OBJECTIVE: Variable methods of animal model have been introduced to develop tracheal stenosis. However, none of the prior models allow for predictable determination of the grade of stenosis. This study sought to establish an animal model of tracheal stenosis by using a cylindrical diffuser and to evaluate the feasibility of a reproducible model. STUDY DESIGN/MATERIALS AND METHODS: A cylindrical diffuser was developed to have a 5 mm active segment to emit laser light circumferentially. Twenty one New Zealand white rabbits were enrolled in this study. The cylindrical diffuser was inserted transorally under bronchoscopic view and the diffused light was delivered to tracheal mucosa 2 cm below the level of vocal cord. Input power of irradiation was 10 W, 5 seconds in group A (n = 7), 10 W, 7 seconds in group B (n = 7), and 8 W, 5 seconds in group C (n = 7). The degree of tracheal stenosis was observed weekly and the rabbits were euthanized 4 weeks after the laser irradiation. RESULTS: The degree of stenosis in group B (90-98%) was significantly larger than that of group A (75-92%) (P = 0.004), while degree in group C (24-35%) was significantly smaller than that of group A (P < 0.001). Two rabbits of group A were euthanized at 3 weeks due to costal retraction. In group B, six rabbits died within 3 weeks after laser irradiation due to severe tracheal stenosis and tracheal malacia, while one rabbit was euthanized 16 days after the irradiation. All rabbits in group C survived up to 4 weeks. Survival between three groups showed significant difference (P = 0.001). CONCLUSION: The degree of stenosis was significantly different according to the delivered optical energy to tracheal mucosa. Therefore, the proposed model may be used in animal studies to emulate variable grades of tracheal stenosis. Lasers Surg. Med. 49:372-379, 2017. © 2016 Wiley Periodicals, Inc.

[Tracheal Surgery]. / Chirurgie der Trachea.

Surgery of the trachea is a specialised field in which many disciplines work jointly due to the variety of indications and the extended topography. Not only because of its particular functional importance, but also because of its complex morphology, anatomy and physiology, this organ represents a special therapeutic challenge. A variety of diseases require surgical procedures of the trachea; the therapeutic strategy is influenced both by the disease itself as well as patient-dependent parameters. Regardless of the nature of the underlying disorder, good results require a high level of expertise in airway management, a careful diagnosis and interventional planning as well as an experienced surgical team that masters extended operative techniques. An optimal treatment decision always requires a multidisciplinary assessment of the patient's individual situation by interventional pulmonologists, thoracic surgeons, visceral surgeons, ENT (ear, nose and throat) surgeons and anaesthesiologists.

Inhibitory Effect of ß-Elemene on Human Airway Granulation Tissue in vivo and in vitro.

BACKGROUND: Recurrent airway granulation hyperplasia and scar formation make airway stenosis a clinical challenge. Therefore, a new approach for the treatment of airway stenosis is necessary. OBJECTIVE: To explore the inhibitory effect of ß-elemene on the proliferation of fibroblasts and airway granulation. METHODS: In vivo: (1) study of the effect of local ß-elemene injection by bronchoscopy. (2) During bronchoscopy, granulation tissues both before and after treatment were obtained. HE staining was performed and the result compared. In vitro: (1) human airway primary fibroblasts were purified and characterized. (2) Fibroblasts were treated with ß-elemene and normal saline (NS) and then examined by optical and electron microscopy. (3) Fibroblasts treated with ß-elemene or NS were assessed for viability by tetrazolium salt assay. (4) Apoptotic rates were determined by flow cytometry. RESULTS: In vivo: (1) after local injection of ß- elemene, airway granulation tissue was reduced. (2) Granulation tissue was found to have less edema, and fibroblasts turned into mature fiber cells. In vitro: (1) human airway primary fibroblasts were successfully purified and cultured. (2) Compared with the control group, fibroblasts of the experimental group became clumped, the plasma granules were increased, and some fibroblasts lost their nucleus and organelles. (3) Compared with the control group, reduction of cell viability was detected with increased concentrations of ß-elemene. (4) With increased concentrations of ß-elemene, apoptotic rates of the fibroblasts were raised compared with the control group. CONCLUSIONS: ß-Elemene may induce apoptosis and necrosis of airway primary fibroblasts and inhibit the proliferation of fibroblasts and airway granulation. The results provide a new approach for the treatment of airway stenosis.

Morgagni hernia presenting with massive pericardial effusion and ascites: prenatal management by thoraco-amniotic shunting and fetal endoscopic tracheal occlusion (FETO) and review of the literature.

BACKGROUND: Morgagni hernia presents a rare type of congenital diaphragmatic hernia (CDH, about 2-5 %) (Herman, J Perinatol 21:343-344, 2001), which is characterized by an anterior mainly right-sided defect of the diaphragm. Infrequently, this is combined with a herniation of the liver into the pericardial cavity (Aké, Prenat Diagn 11:719-724, 1991; Stevens, Pediatr Radiol 26:791-793, 1996). This may cause massive pericardial effusion and subsequently lung hypoplasia (Pober et al., Congenital diaphragmatic hernia overview, University of Washington, Seattle, 2015; Ikeda, J Perinat Med 30:336-340, 2002; Hara, J Obstet Gynaecol Res 33:561-565, 2007). So far only few cases have been reported in fetal life. CASE: We report a case of Morgagni hernia with pericardiodiaphragmatic aplasia, complicated by two-compartment effusions (massive pericardial effusion and mild ascites), diagnosed in the second trimester. The case was successfully managed in utero with thoraco-amniotic shunting and late tracheal occlusion, followed by corrective surgery after birth. DISCUSSION: A review of the literature was performed, identifying 13 cases of prenatally diagnosed Morgagni hernia. The diagnosis was established by the sonographic findings of pericardial effusion und intrathoracic herniation of the liver. In only two cases a prenatal intervention was carried out. All neonates were operated postnatally with excellent final outcome.

Frontometaphyseal dysplasia and keloid formation without FLNA mutations.

Frontometaphyseal dysplasia (FMD) is a distinctive sclerosing skeletal dysplasia associated with a number of non-skeletal manifestations including hearing loss, cardiac malformations, and stenosis, particularly of the upper airway and urinary tract. Some, but not all, patients have mutations in FLNA causing the condition. Consonant with the X chromosomal location of FLNA males are generally more severely affected than females. FLNA mutations can be detected in 82% of affected males. We describe seven patients (one male, six females) all of whom have the major clinical and radiological features of FMD, but without detectable mutations in FLNA. The females in our cohort are affected to a similar degree as is usually found in males. In addition, all patients have marked keloid formation at various body sites, including the eye, from an early age. Other features that may indicate a different etiology in these patients are the increased frequency of cleft palate, Robin sequence, tracheal stenosis, and mild intellectual disability, which all occur in three of more patients in the present group. All patients are isolated. We hypothesize that the presently reported patients represent further evidence that phenotypes strongly resembling FMD exist that are not accounted for by mutations in FLNA. Since the frequency of several of the manifestations, their sporadic presentations, and the presence of keloid formation differ from the X-linked form of this condition we propose de novo autosomal dominant acting mutations in a gene functionally related to FLNA, underpin this disorder.

Influence of Tracheal Obstruction on the Efficacy of Superimposed High-frequency Jet Ventilation and Single-frequency Jet Ventilation.

BACKGROUND: Both superimposed high-frequency jet ventilation (SHFJV) and single-frequency (high-frequency) jet ventilation (HFJV) have been used with success for airway surgery, but SHFJV has been found to provide higher lung volumes and better gas exchange than HFJV in unobstructed airways. The authors systematically compared the ventilation efficacy of SHFJV and HFJV at different ventilation frequencies in a model of tracheal obstruction and describe the frequency and obstruction dependence of SHFJV efficacy. METHODS: Ten anesthetized animals (weight 25 to 31.5 kg) were alternately ventilated with SHFJV and HFJV at a set of different fHF from 50 to 600 min. Obstruction was created by insertion of interchangeable stents with ID 2 to 8 mm into the trachea. Chest wall volume was measured using optoelectronic plethysmography, airway pressures were recorded, and blood gases were analyzed repeatedly. RESULTS: SHFJV provided greater than 1.6 times higher end-expiratory chest wall volume than HFJV, and tidal volume (VT) was always greater than 200 ml with SHFJV. Increase of fHF from 50 to 600 min during HFJV resulted in a more than 30-fold VT decrease from 112 ml (97 to 130 ml) to negligible values and resulted in severe hypoxia and hypercapnia. During SHFJV, stent ID reduction from 8 to 2 mm increased end-expiratory chest wall volume by up to 3 times from approximately 100 to 300 ml and decreased VT by up to 4.2 times from approximately 470 to 110 ml. Oxygenation and ventilation were acceptable for 4 mm ID or more, but hypercapnia occurred with the 2 mm stent. CONCLUSION: In this in vivo porcine model of variable severe tracheal stenosis, SHFJV effectively increased lung volumes and maintained gas exchange and may be advantageous in severe airway obstruction.

Multilevel airway stenosis in patients with granulomatosis with polyangiitis (Wegener's).

OBJECTIVES: To describe the presentation and clinical course of subglottic stenosis (SGS), in particular the development of concurrent airway lesions, in patients with Granulomatosis with Polyangiitis (Wegener's) (GPA). MATERIALS AND METHODS: Retrospective review of clinical data from all patients presenting to our institution from 2000 to 2012 with SGS and GPA. RESULTS: Thirty-five patients were identified. The average age at diagnosis was 33 years old. Eleven patients (31%) presented with SGS as part their initial manifestation of GPA. The remaining patients developed SGS later, at a median of 2.5 years from diagnosis (range 6 months to 14 years). Twelve patients (34%) were noted to have multilevel airway involvement. Seven patients (20%) had documentation of cricoarytenoid joint fixation and vocal cord immobility. This was typically progressive in nature and occurred at an average of two years following the diagnosis of SGS. Six patients (17%) had mid/distal tracheal stenosis and four (11%) had bronchial stenosis. The majority of patients (86%) had evidence of concurrent sinonasal involvement, ten patients (29%) had evidence of otologic involvement and eight (23%) had ocular involvement. CONCLUSIONS: Cricoarytenoid joint fixation and distal stenosis occur not infrequently in patients with GPA and SGS, resulting in progressive multilevel airway stenosis in about one third of patients. It is critical to identify multilevel stenosis when managing the airways of these patients.

COMPARISON OF THE RADIOGRAPHIC AND TRACHEOSCOPIC APPEARANCE OF THE DORSAL TRACHEAL MEMBRANE IN LARGE AND SMALL BREED DOGS.

The etiology and clinical significance of increased radiographic opacity along the dorsal margin of the tracheal lumen has long been debated. Most often, this opacity is attributed to redundancy of the dorsal tracheal membrane (DTM), a condition that occurs with tracheal collapse. We hypothesized that the underlying etiology of this radiographic opacity differs between small breed dogs with tracheal collapse and small or large breed dogs without tracheal collapse. The purpose of this prospective, cross-sectional study was to compare the radiographic appearance of an increased opacity within the trachea to tracheoscopy findings in a group of small and large breed dogs. A total of 17 small breed dogs and 16 large breed dogs were included. Of these, only one did not have a radiographically visible DTM. Small breed dogs were divided into groups with tracheal collapse (n = 8) and those without (n = 9) based on tracheoscopy. Tracheal collapse was absent in larger breed dogs, however both large and small breed dogs demonstrated inward invagination of the DTM. In dogs with tracheal collapse, the DTM occupied a larger percentage of the tracheal luminal height on radiographs and a larger percentage of tracheal circumference on tracheoscopy vs. dogs with an invaginated DTM on tracheoscopy and dogs with no collapse and no invagination of the DTM. Findings supported the hypothesis that increased radiographic opacity along the dorsal margin of the trachea arises from different etiologies in dogs with and without tracheal collapse.

Use of a Nitinol Wire Stent for Management of Severe Tracheal Stenosis in an Eclectus Parrot (Eclectus roratus).

A 25-year-old, female eclectus parrot (Eclectus roratus) presented for dyspnea 3 weeks after anesthesia and surgery for egg yolk coelomitis. Radiography, computed tomography, and tracheoscopy revealed multiple tracheal strictures spanning a length of 2.6 cm in the mid to distal trachea. Histopathologic examination revealed mild fibrosis, inflammation, and hyperplasia consistent with acquired tracheal strictures. Tracheal resection was not considered possible because of the length of the affected trachea. The strictures were resected endoscopically, and repeated balloon dilation under fluoroscopic guidance over the course of 10 months resulted in immediate but unsustained improvement. Computed tomography was used to measure the stenotic area. A 4 × 36-mm, custom-made, nitinol wire stent was inserted into the trachea under fluoroscopic guidance. After stent placement, intermittent episodes of mild to moderate dyspnea continued, and these responded to nebulization with a combination of saline, acetylcysteine, and dexamethasone. Multiple attempts to wean the patient off nebulization therapy and to switch to a corticosteroid-free combination were unsuccessful. The parrot eventually developed complications, was euthanatized, and necropsy was performed. Histologically, the tracheal mucosa had widespread erosion to ulceration, with accumulation of intraluminal exudate and bacteria, severe degeneration of skeletal muscle and tracheal rings, prominent fibrosis, and mild to moderate, submucosal inflammation. Clinicopathologic findings in this case suggested tracheomalacia, which has not been previously described in birds. Custom-made tracheal stents can be used for severe tracheal stenosis in birds when tracheal resection and anastomosis is not possible. Complications of tracheal stent placement in birds may include tracheitis and tracheomalacia. To our knowledge, this is the first report of tracheal stent placement in an avian species.

Microscopic examination of iatrogenic subglottic tracheal stenosis: observations that may elucidate its histopathologic origin.

OBJECTIVES: The histopathologic origin of iatrogenic subglottic tracheal stenosis (ISTS) remains unclear. The purpose of this study was to use detailed operative microscopy to systematically examine the operative en bloc specimens of patients with ISTS and to observe the histologic and morphological changes in the hopes that these observations will provide insight into the histopathologic origin of these devastating injuries. METHODS: The operative specimens of 18 patients who underwent open tracheal or laryngotracheal resection for ISTS were examined. Precise morphological characteristics were investigated for each tissue layer, including the adventitia, the outer surface of the perichondrium, the cartilage, the inner surface of the perichondrium, the submucosa, and the mucosa. Each tissue layer was evaluated independently and in relationship to the other layers. The cartilaginous airway was further evaluated relative to the pars membranacea. RESULTS: The most common morphological finding in the epithelium was squamous metaplasia with occasional intense inflammation visible in the underlying mucosa, including cicatrization. The underlying cartilage demonstrated ossific metaplasia with sequestration in many cases. By far the most pronounced changes were found in the outer perichondrium and overlying adventitia and included diffuse paucicellular or hyperplastic fibrosis with intense hyperplastic scar formation or hyaline cicatrization. In the pars membranacea, severe scar formation and hyperplastic fibrosis were predominant. Ossific metaplasia was particularly severe in the lateral or outer parts of the tracheal ring, particularly in the vicinity of the adventitia and outer perichondrium. These changes were much more pronounced than the relatively minor changes observed in the submucosa and mucosa. CONCLUSIONS: The most severe pathologic observations occurred in the lateral tissue layers, ie, the outer perichondrium and adventia. Given that an injury occurs from the tracheal lumen, these tissue layers have the greatest distance from the site of injury. As only minor changes occurred in the inner tissue layers, we hypothesize that these tissues have a greater regenerative capacity than the outer layers. This study supports the theory that the depth of the airway injury is more critical to the development of ISTS than is the extent or length of the injury.

Airway management and endoscopic treatment of subglottic and tracheal stenosis: the laryngeal mask airway technique.

OBJECTIVES: The objective is to present clinical outcomes of subglottic and tracheal stenosis treated by flexible bronchoscopic delivery of carbon dioxide (CO2) laser via laryngeal mask airway (LMA). METHODS: All consecutive, nontracheotomy dependent cases of subglottic and tracheal stenosis treated endoscopically over a 4-year period were retrospectively reviewed. The surgical approach consisted of radial incisions using a flexible fiber-based CO2 laser, balloon dilation, and topical application of mitomycin C. Ventilation during the procedure occurred through the LMA, and the CO2 laser fiber was delivered through the working channel of a flexible bronchoscope passed through the LMA. Number of dilations, period between dilations, and operative times were reviewed. RESULTS: Eleven patients who underwent airway intervention during the study period were identified. Average follow-up was 28 months. Etiologies of airway stenosis included intubation injury (6), idiopathic (4), or autoimmune disease (1), requiring an average of 1.3, 1.5, and 3 dilations, respectively. Average operative time was 67 minutes. Autoimmune etiology correlated with more frequent dilations. CONCLUSION: LMA is an effective way to manage ventilation while simultaneously allowing unencumbered flexible bronchoscopic access for laser surgery, balloon dilation, and mitomycin C application for airway stenosis. Long-term success in treating stenosis is achievable using this technique.

Finding balance between minimally invasive surgery and laryngotracheal resection in the management of adult laryngotracheal stenosis.

Management of adult laryngotracheal stenosis is complex and several treatment options are known. The present study focuses on finding the right balance between minimally invasive surgery and laryngotracheal resection by reviewing a single institution's experiences. Retrospective analysis was performed of all adult and adolescent patients with laryngotracheal stenosis who underwent treatment in a tertiary referral center, between 1990 and 2012. Age, gender, etiology, treatment, recurrence, pre- and post-operative peak flow (PF), and pre- and post-treatment subjective complain scores (SCS) were registered. 87 patients with 267 interventions were analyzed. There were 238 dilatation tracheoscopies, 22 open surgeries and various other endoscopic procedures registered. Idiopathic stenoses required the most dilatation tracheoscopies, while post-tracheotomy stenoses required the least. Patients in the post-intubation and post-tracheotomy groups were significantly more often treated with open surgery compared to those in the granulomatosis with polyangiitis (GPA) and idiopathic groups. The gain in PF flow after dilatation tracheoscopy was significantly higher in the idiopathic group compared to the other groups. The median SCS of dyspnoea decreased in the whole population, while other SCS did not change remarkably. Repeated endoscopic procedures are recommended in patients with severe systemic disease which do not allow open surgery or when other comorbidities contraindicate open surgery. Open surgery very often offers the definitive solution in the treatment of laryngotracheal stenosis and cannot be avoided when the laryngeal or the tracheal framework is damaged. Patients' personal preferences have to be considered in the pre-operative assessment process.