Pleurodesis with Thulium Cyber Laser versus talc poudrage: a comparative experimental study.

Sclerosing fluids to achieve pleurodesis could be hardly replaced for bed-side procedures, but other devices may be successfully applied during thoracoscopy. Thulium Cyber Laser was experimented for this purpose and compared to talc poudrage. Twenty pigs underwent operative videothoracoscopy (VATS). Ten models were subjected to double-port VATS and parietal pleura photoevaporation using Thulium Cyber Laser™ (TCL) 150 W 2010 nm on the posterior third of three ribs; the pleural surface was homogeneously treated inside the target perimeter. The remaining ten pigs underwent uniportal thoracoscopy; talc poudrage was performed using the current clinical practice dosage (1 g/18 kg) with accurate talc powder spread over the whole pleural surface. All models were followed up for 60 days. Pleurodesis firmness was graded on a three-tier scale (none-moderate-firm) and site-matching topographical expectancy was evaluated. TCL produced pleurodesis in all models: 7/10 were firm and 3/10 moderate. Talc poudrage pleurodesis was firm in 4/10 and moderate in 6/10. Pleural adhesions were found exclusively in the treated area after laser treatment, while talc created a wide spectrum of effects, most commonly anarchic jagged adhesions obliterating less than 50 % of the pleural cavity (7/10), mostly declivous. The pathologist found more aggressive inflammation (sometimes severe) in the talc group. Expected localized pleurodesis was always registered in laser group (10/10), while talc poudrage was found poorly effective if consistent pleurodesis is expected in an apico-dorsal position (2/10). Laser pleurodesis appears more homogeneous, qualitatively not inferior, and topographically more predictable than talc pleurodesis. Parietal photoevaporation seems effective and the localized pleurodesis is reproducible.

An Expectation-Maximization Algorithm for Including Oncological COVID-19 Deaths in Survival Analysis.

We address the problem of how COVID-19 deaths observed in an oncology clinical trial can be consistently taken into account in typical survival estimates. We refer to oncological patients since there is empirical evidence of strong correlation between COVID-19 and cancer deaths, which implies that COVID-19 deaths cannot be treated simply as non-informative censoring, a property usually required by the classical survival estimators. We consider the problem in the framework of the widely used Kaplan-Meier (KM) estimator. Through a counterfactual approach, an algorithmic method is developed allowing to include COVID-19 deaths in the observed data by mean-imputation. The procedure can be seen in the class of the Expectation-Maximization (EM) algorithms and will be referred to as Covid-Death Mean-Imputation (CoDMI) algorithm. We discuss the CoDMI underlying assumptions and the convergence issue. The algorithm provides a completed lifetime data set, where each Covid-death time is replaced by a point estimate of the corresponding virtual lifetime. This complete data set is naturally equipped with the corresponding KM survival function estimate and all available statistical tools can be applied to these data. However, mean-imputation requires an increased variance of the estimates. We then propose a natural extension of the classical Greenwood's formula, thus obtaining expanded confidence intervals for the survival function estimate. To illustrate how the algorithm works, CoDMI is applied to real medical data extended by the addition of artificial Covid-death observations. The results are compared with the estimates provided by the two naïve approaches which count COVID-19 deaths as censoring or as deaths by the disease under study. In order to evaluate the predictive performances of CoDMI an extensive simulation study is carried out. The results indicate that in the simulated scenarios CoDMI is roughly unbiased and outperforms the estimates obtained by the naïve approaches. A user-friendly version of CoDMI programmed in R is freely available.

Survival Analysis Using the Covid-death Mean-imputation (CoDMI) Algorithm: A First Clinical Application in Radiation Oncology.

BACKGROUND/AIM: To report long-term survival results after trimodal approach for locally advanced rectal cancer (LARC) in the Covid-19 era. We herein illustrate a clinical application of Covid-death mean-imputation (CoDMI) algorithm in LARC patients with Covid-19 infection. PATIENTS AND METHODS: We analyzed 94 patients treated for primary LARC. Overall survival was calculated in months from diagnosis to first event (last follow-up/death). Because Covid-19 death events potentially bias survival estimation, to eliminate skewed data due to Covid-19 death events, the observed lifetime of Covid-19 cases was replaced by its corresponding expected lifetime in absence of the Covid-19 event using the CoDMI algorithm. Patients who died of Covid-19 (DoC) are mean-imputed by the Kaplan-Meier estimator. Under this approach, the observed lifetime of each DoC patient is considered as an "incomplete data" and is extended by an additional expected lifetime computed using the classical Kaplan-Meier model. RESULTS: Sixteen patients were dead of disease (DoD), 1 patient was DoC and 77 cases were censored (Cen). The DoC patient died of Covid-19 52 months after diagnosis. The CoDMI algorithm computed the expected future lifetime provided by the Kaplan-Meier estimator applied to the no-DoC observations as well as to the DoC data itself. Given the DoC event at 52 months, the CoDMI algorithm estimated that this patient would have died after 79.5 months of follow-up. CONCLUSION: The CoDMI algorithm leads to "unbiased" probability of overall survival in LARC patients with Covid-19 infection, compared to that provided by a naïve application of Kaplan-Meier approach. This allows for a proper interpretation/use of Covid-19 events in survival analysis. A user-friendly version of CoDMI is freely available at https://github.com/alef-innovation/codmi.

Equine Adipose-Derived Mesenchymal Stromal Cells Release Extracellular Vesicles Enclosing Different Subsets of Small RNAs.

BACKGROUND: Equine adipose-derived mesenchymal stromal cells (e-AdMSC) exhibit attractive proregenerative properties strongly related to the delivery of extracellular vesicles (EVs) that enclose different kinds of molecules including RNAs. In this study, we investigated small RNA content of EVs produced by e-AdMSC with the aim of speculating on their possible biological role. METHODS: EVs were obtained by ultracentrifugation of the conditioned medium of e-AdMSC of 4 subjects. Transmission electron microscopy and scanning electron microscopy were performed to assess their size and nanostructure. RNA was isolated, enriched for small RNAs (<200 nt), and sequenced by Illumina technology. After bioinformatic analysis with state-of-the-art pipelines for short sequences, mapped reads were used to describe EV RNA cargo, reporting classes, and abundances. Enrichment analyses were performed to infer involved pathways and functional categories. RESULTS: Electron microscopy showed the presence of vesicles ranging in size from 30 to 300 nm and expressing typical markers. RNA analysis revealed that ribosomal RNA was the most abundant fraction, followed by small nucleolar RNAs (snoRNAs, 13.67%). Miscellaneous RNA (misc_RNA) reached 4.57% of the total where Y RNA, RNaseP, and vault RNA represented the main categories. miRNAs were sequenced at a lower level (3.51%) as well as protein-coding genes (1.33%). Pathway analyses on the protein-coding fraction revealed a significant enrichment for the "ribosome" pathway followed by "oxidative phosphorylation." Gene Ontology analysis showed enrichment for terms like "extracellular exosome," "organelle envelope," "RNA binding," and "small molecule metabolic process." The miRNA target pathway analysis revealed the presence of "signaling pathways regulating pluripotency of stem cells" coherent with the source of the samples. CONCLUSION: We herein demonstrated that e-AdMSC release EVs enclosing different subsets of small RNAs that potentially regulate a number of biological processes. These findings shed light on the role of EVs in the context of MSC biology.

Effectiveness and predictability of pleurodesis with the Tachosil® surgical patch compared with talc poudrage: an experimental study.

OBJECTIVES: The human fibrinogen/thrombin surgical patch is proposed for videothoracoscopic pleurodesis after recurrent observation in routine clinical practice of site-specific pleuro-pulmonary adhesions. METHODS: A total of 36 experimental videothoracoscopies were performed on 18 pigs, all of which underwent a bilateral sequential procedure. The right or left side was casually approached and two 4.8 × 4.8 cm Tachosil® patches were placed over the dorsal lung surface in the highest antigravitary position through a double-port thoracoscopy. The contralateral side was subsequently entered by a single-port thoracoscopy to perform talc poudrage in accordance with current recommendations (1 g/18 kg). Thoracoscopy and autopsy were performed after 60 days to evaluate the outcome. Pleurodesis was graded on a predefined three-tier scale (none-moderate-firm); specimens were then submitted to the pathologist. The statistical analyses performed were: frequencies distribution, variables association test and hypothesis testing using binomial distribution. RESULTS: Pleurodesis was firm in 15/18 cases (P = 0.004), moderate in 2/18, and none in 1/18 cases with the sealant matrix. Talc pleurodesis was firm in 7/18 cases and moderate in 11/18. No surgical, medical, behavioural or growth disorders were recorded. There was complete correspondence between lung surface covered by the sealant patch and pleural adhesion topography (P < 0.001) and no other sites of adhesion were found at autopsy, whereas talc effect was commonly found to produce random pleural cavity obliteration, most frequently in declivous places, with inhomogeneous pleural adhesion for tissue density and rough/smooth outline (P < 0.001). The pathologist disclosed that talc creates a variable and unpredictable inflammatory background, sometimes severe, with the development of many granulomas. Fibrin sealant patch pleurodesis is caused by connective tissue irrespective of the pleural and pulmonary anatomy. CONCLUSIONS: The fibrin sealant patch is effective in producing localized pleurodesis in swine. The morphology and topography of the adhesions, 2 months after the patch placement, are consistent and more predictable than after talc poudrage. The latter causes a diffuse pleurodesis which is not predictable, with a predominantly gravitational location and produces a much more serious inflammatory response.

Gambling on putative biomarkers of osteoarthritis and osteochondrosis by equine synovial fluid proteomics.

Osteoarthritis (OA) and osteochondrosis (OC) are two of the main challenges in orthopedics, whose definitive diagnosis is usually based on radiographic/arthroscopic evidences. Their early diagnosis should allow preventive or timely therapeutic actions, which are generally precluded from the poor relationships occurring between symptomatologic and radiographic evidences. These limitations should be overcome by improving the knowledge on articular tissue metabolism and on molecular factors regulating its normal homeostasis, also identifying novel OA and OC biomarkers suitable for their earlier diagnoses, whenever clinical/pathological inflammatory scenarios between these joint diseases seem somewhat related. To identify proteins involved in their aetiology and progression, we undertook a differential proteomic analysis of equine synovial fluid (SF), which compared the protein pattern of OA or OC patients with that of healthy individuals. Deregulated proteins in OA and OC included components related to inflammatory state, coagulation pathways, oxidative stress and matrix damage, which were suggestive of pathological alterations in articular homeostasis, plasma-SF exchange, joint nutritional status and vessel permeability. Some proteins seemed commonly deregulated in both pathologies indicating that, regardless of the stimulus, common pathways are affected and/or the animal joint uses the same molecular mechanisms to restore its homeostasis. On the other hand, the increased number of deregulated proteins observed in OA with respect to OC, together with their nature, confirmed the high inflammatory character of this disease. Some deregulated proteins in OA found a verification by analyzing the SF of injured arthritic joints following autologous conditioned serum treatment, an emergent therapy that provides positive results for both human and equine OA. Being the horse involved in occupational/sporting activities and considered as an excellent animal model for human joint diseases, our data provide suggestive information for tentative biomedical extrapolations, allowing to overcome the limitations in joint size and workload that are typical of other small animal models.

Radiographic evaluation of changes in the proximal phalanx of Thoroughbreds in race training.

OBJECTIVE: To evaluate changes in the cortical bone of the proximal phalanx of the fore-limbs of Thoroughbreds in response to training. ANIMALS: Twenty-seven 2-year-old Thoroughbreds (20 females, 2 males, and 5 geldings). PROCEDURES: Horses were principally in training for races in a straight line and in a clockwise direction. Lateromedial and dorsopalmar radiographic views of each metacarpophalangeal joint were obtained before the horses started training and 1 year after starting exercise and racing. Width of the dorsal, palmar, lateral, and medial cortex and the width and thickness of the medulla were measured. Ratios (rather than absolute values) were used to remove the effect of differences in bone size among horses. RESULTS: 10 horses were lost from the study. Radiographs were obtained for 17 horses 1 year after starting training (9 horses raced in a clockwise direction, and 8 raced in clockwise and counterclockwise directions). There was no difference between the cortical bone in the right and left forelimbs at the start of the study. After training for 1 year, the palmar cortex in the right forelimb was significantly thicker than that in the left forelimb. CONCLUSIONS AND CLINICAL RELEVANCE: The strain patterns, biomechanics of rapid exercise, and type of training most probably determined differences in the adaptive responses of the proximal phalanx. The data reported here can be used in the evaluation of weight-bearing distribution along the proximal phalanx and evaluation of the relationship between exercise and bone remodelling of the proximal phalanx.

Laparoscopic sterilization of Sardinia donkeys using an endoscopic stapler.

OBJECTIVE: To describe a laparoscopic technique for sterilization of the male donkey using an endoscopic linear stapler (ELS). STUDY DESIGN: Clinical report. ANIMALS: Male Sardinia donkeys (n=6), aged 24-36 months. METHODS: Laparoscopic sterilization was performed under standing sedation or recumbent under general anesthesia. An endoscopic linear stapler was used to transect, and control hemorrhage of, the spermatic cord. Serum testosterone concentrations were measured preoperatively and at 3, 6, and 12 months after sterilization. One testis from each of 2 donkeys was collected at 12 months for histologic examination. RESULTS: Sterilization was successfully achieved. The only minor intraoperative complication experienced was hemorrhage from 1 spermatic cord stump, which was re-stapled. No other major short- or long-term complications occurred. Serum testosterone concentrations were < 15 ng/dL at 3, 6, and 12 months after surgery in all donkeys. Testicular atrophy without evidence of revascularization was observed in the 2 testes examined histologically. CONCLUSIONS: Sterilization of descended testes by intraabdominal laparoscopic transection of the spermatic cord is a simple practical method, with minimal intraoperative or postoperative complications. Testicular revascularization did not occur. CLINICAL RELEVANCE: Laparoscopic sterilization in donkey can be performed using an endoscopic linear stapler as an alternative to scrotal castration.