Production and concentration of keratinases and application of fermentation residual in removing hexavalent chromium.

The production of keratinases was evaluated in submerged fermentation with Aspergillus niger and by pigs' swine hair in a batch bioreactor. Experimental planning was performed to assess the interaction between different variables. The enzyme extract produced was characterized at various pH and temperatures and subjected to enzyme concentration using a biphasic aqueous system and salt/solvent precipitation techniques. In addition, the substrate's potential in reducing hexavalent chromium from synthetic potassium dichromate effluent with an initial concentration of 20 mg L-1 of chromium was evaluated. The resulting enzyme extract showed 89 ± 2 U mL-1 of keratinase. The enzyme concentration resulted in a purification factor of 1.3, while sodium chloride/acetone and ammonium sulfate/acetone resulted in a purification factor of 1.9 and 1.4, respectively. Still using the residual substrate of swine hair from the fermentation, a 94% reduction of hexavalent chromium concentration occurred after 9 h of reaction. Thus, the study proved relevant for producing keratinases, with further environmental applicability and the possibility of concentrating the extract via low-cost processes.

Study and characterization of a product based on a vegetable extract of quinoa fermented with water kefir grains.

This work aimed to study and characterize a product based on vegetable extract of quinoa (WVEQ) fermented with water kefir grains. The effect of sucrose concentration (SC), inulin concentration (IC), and xanthan gum (XG) concentration were evaluated using a central composite design (CCD) 23. They were subsequently characterized regarding cellular growth of the grains, beverage yield, pH, soluble solids, carbon dioxide (CO2) production, lactic acid, and ethanol production. Therefore, for the final stage, two formulations (F1 and F8) of the CCD were chosen to be characterized in terms of proximate composition, microbiological composition of the kefir culture, analysis of organic compounds, sensory analysis, and enzymatic and microbiological characterization before and after simulation of in vitro gastrointestinal digestion. In the two chosen products, one can see that fermentation optimized the bioavailability of proteins due to the high proteolytic activity of the microorganisms in kefir and the increase in lipid content. In identifying microorganisms, there was a prevalence of Saccharomyces sp. yeasts. In the sensory analysis, the F8 formulation showed better results than the F1 formulation. In vitro, gastrointestinal digestion showed reduced lactic acid bacteria and yeast and increased acetic acid bacteria in the liquid phase for both formulations. In the enzymatic profile, there was a reduction in all enzymes analyzed for both formulations, except for amylase in F1, which went from 14.05 U/mL to 39.41 U/mL. Therefore, it is concluded that using WVEQ as a substrate for the product appears to be a viable alternative with nutritional and technological advantages for serving a specific market niche.

Trichoderma koningiopsis fermentation in airlift bioreactor for bioherbicide production.

During scaling of fermentations, choosing a bioreactor is fundamental to ensure the product's quality. This study aims to produce bioherbicides using Trichoderma koningiopsis fermentation, evaluating process parameters in an Airlift bioreactor. As a response, we quantified the production of enzymes involved in the bioherbicide activity (amylase, cellulase, laccase, lipase, and peroxidase). In addition, it evaluated the agronomic efficiency of the fermented extract optimized through tests that promoted soybean growth and nodulation, soybean seed germination, and in vitro phytopathogen control. As a result of optimizing the scaling bioprocess, it was possible to obtain an adequate fermentation condition, which, when applied to soybean seeds, had beneficial effects on their growth. It allowed the production of an enzyme cocktail. These results add a crucial biotechnological potential factor for the success of the optimized formulation in the Airlift bioreactor, in addition to presenting relevant results for the scientific community.

Six-year Burden of Care for Nonsyndromic Unilateral Cleft Lip and Palate Patients: A Comparison Between Cleft Centers and Noncleft Centers.

OBJECTIVE: To determine differences in burden of care between nonsyndromic patients with unilateral cleft lip and palate undergoing treatment at American Cleft Palate-Craniofacial Association (ACPA)-accredited centers and nonaccredited centers in New York State. DESIGN: A retrospective review of the New York Statewide Planning and Research Cooperative System database from January 2001 to December 2014 was performed using ICD-9 and CPT coding. PATIENTS, PARTICIPANTS: This study included patients with unilateral cleft lip and palate who underwent both lip and palate repairs during their first 6 years of life. Exclusion criteria included orofacial cleft syndromes, follow-up under 6 years, and one-stage combined cleft lip and palate repairs. RESULTS: Eighty-eight patients were treated at cleft centers, and 29 patients at nonaccredited centers ( n = 117). Age at primary palatoplasty (13.0 months vs 18.1 months; p = .019), total number of cleft operations (2.3 vs 2.7; p = .012), and total number of primary cleft-specific procedures (2.2 vs 2.5; p = .0049) were significantly lower for patients treated in cleft centers. Age at primary cheiloplasty (4.8 months vs 4.6 months; p = .865), post-cheiloplasty length of stay (1.2 days vs 1.2 days; p = .673), post-palatoplasty length of stay (1.5 days vs 1.9 days; p = .211), average hospital admissions (2.2 vs 2.3; p = 0.161), and total complication rates (34.1% vs 21.1%; p = 0.517) did not differ significantly between cleft centers and noncenters. CONCLUSIONS: This data demonstrates some significant differences in overall 6 year burden of care for nonsyndromic patients with unilateral cleft lip and palate treated at ACPA-accredited cleft centers versus nonaccredited centers.

The hepatic extramedullary hematopoiesis during experimental murine Schistosomiasis mansoni.

Many years ago, our research group has demonstrated extramedullary hematopoiesis in the peripheral zone of murine hepatic schistosomal granulomas. In the present study, we revisit this phenomenon using new technical and conceptual approaches. Therefore, newborn mice were percutaneously infected by Schistosoma mansoni cercariae and euthanized between 35- and 60-days post infection. Liver samples were submitted to histopathology and immunohistochemical analyses. Cells under mitosis and/or expressing Ki67 demonstrated the proliferation of hematopoietic cells both around the parasite's eggs trapped in the liver and around hepatic vessels. After 50 days post infection, proliferating cells at different levels on differentiation were located preferentially in the peripheral zone of the granulomas, around the vessels and inside the sinusoids. The presence of acidic and sulfated glycoconjugates, reticular fibers and the absence of fibronectin characterized the microenvironment for attraction and maintenance of hematopoiesis. Some neutrophils secreted MMP9 from the earliest points of infection, indicating degradation of the extracellular matrix in regions of histolysis and a possible chemoattraction of hematopoietic stem cells to the liver. Fall-3+ cells and Sca-1+ cells indicated that early hematopoietic progenitors could be mobilized to the liver. Groups of vWF+ megakaryocytes suggest chemoattraction of these cells and/or migration, proliferation, and differentiation of very immature progenitors to this organ. The increase of blood vessels and extramedullary hematopoiesis in this environment, where markers of immature hematopoietic and endothelial cells have been identified, points to the possibility of the presence of progenitors for endothelial and hematopoietic cells in the liver during the infection. There is also the possibility of concomitant migration of more differentiated hematopoietic progenitors, that proliferate and differentiate in the liver, and the occurrence of angiogenesis caused by inflammation or release of ovular antigens that stimulate the activation and proliferation of endothelial cells. Altogether, these data increase knowledge about a murine model that is of interest for investigating the pathology of the schistosomiasis and also the dynamics of hematopoiesis.

Clinical Outcomes After Hematoma Development: A Study of 627 Tissue Expander Breast Reconstructions.

BACKGROUND: Hematomas after tissue expander immediate breast reconstruction (TE-IBR) pose a significant challenge during the recovery period. In this study, we aim to evaluate whether hematoma formation leads to subsequent complications and how management can impact final reconstructive goals. METHODS: A single-institution retrospective review of TE-IBRs from 2001 to 2018 was performed using an established breast reconstruction database. Demographics, medications, comorbidities, and complications were identified. Implant loss was defined as removal of the tissue expander/implant without immediate reimplantation during that operation. Hematoma size, management, transfusion requirement, reoperations, and final outcome were recorded. Reconstructive failure was defined as an implant loss that was not replaced with another implant or required secondary autologous reconstruction. RESULTS: Six hundred twenty-seven TE-IBR patients were analyzed. Postoperative hematoma (group 1) occurred in 4.1% (n = 26) of TE-IBRs and did not develop in 95.9% (group 2: n = 601). Group 2 had a higher mean body mass index (24.5 vs 27.3 kg/m, P = 0.018); however, there were no significant differences in smoking status, preoperative/postoperative radiation/chemotherapy, or other comorbidities. Group 1 was found to have increased rates of implant loss (15.4% vs 3.7%, P = 0.0033) and reconstructive failure (11.5% vs 2.8%, P = 0.0133) compared with group 2.Eighteen hematomas (69.2%) underwent surgical intervention (group 1a) compared with 30.8% (n = 8) that were clinically managed (group 1b). Group 1a had statistically significant lower rates of subsequent complications (22.2% vs 62.5%, P = 0.046) and reoperations (5.6% vs 27.5%, P = 0.037) than did group 1b, respectively.Lastly, 23.1% (n = 6) of patients who developed a hematoma were on home antithrombotics (group 1c) compared with 76.9% (n = 20) of patients with no antithrombotics (group 1d). There were statistically significant differences in transfusion rates (50% vs 0%, P = 0.001) between groups 1c and 1d, respectively. Differences in hematoma volume (330 vs 169.3 mL, P = 0.078) and reconstructive failure (33.3% vs 5%, P = 0.057) approached significance between both groups. CONCLUSIONS: Hematoma after TE-IBR should be monitored closely, as it may play a role in jeopardizing reconstruction success. Patients on home antithrombotic medication may be at increased risk of larger-volume hematomas and reconstruction failure. Plastic surgeons should consider aggressive surgical evacuation of postoperative TE-IBR hematomas to reduce subsequent complications and reoperations, thus optimizing reconstructive outcomes.

Postoperative Venous Thromboembolism Prophylaxis Utilizing Enoxaparin Does Not Increase Bleeding Complications After Abdominal Body Contouring Surgery.

BACKGROUND: Venous thromboembolism (VTE) chemoprophylaxis warrants an individualized, risk-stratified approach, and constitutes a relatively controversial topic in plastic surgery. OBJECTIVES: The aim of this study was to determine the safety of a 7-day postoperative enoxaparin regimen for VTE prophylaxis compared with a single preoperative dose of heparin in abdominal body contouring surgery. METHODS: This single-institution pre-post study investigated the safety of a 7-day enoxaparin postoperative regimen in abdominal body contouring procedures performed by a single surgeon from 2007 to 2018. Four procedures were included: traditional panniculectomy, abdominoplasty, fleur-de-lis panniculectomy, and body contouring liposuction. Group I patients received a single dose of 5000 U subcutaneous heparin in the preoperative period, and no postoperative chemical prophylaxis was administered. Group II patients received 40 mg subcutaneous enoxaparin in the immediate preoperative period, then once daily for 7 days postoperatively. RESULTS: A total of 195 patients were included in the study, 66 in Group I and 129 in Group II. The groups demonstrated statistically similar VTE risk profiles, based on the 2005 Caprini risk-assessment model. There were no statistically significant differences in the 2 primary outcomes: postoperative bleeding and VTE events. Group I patients had higher reoperation rates (22.7% vs 10.1%, P = 0.029), which was secondary to higher rates of revision procedures. CONCLUSIONS: A 7-day postoperative course of once-daily enoxaparin for VTE risk reduction in abdominal body contouring surgery does not significantly increase the risk of bleeding. Implementation of this regimen for postdischarge chemoprophylaxis, when indicated following individualized risk stratification, is appropriate.

Acellular Dermal Matrix Sterility: Does It Affect Microbial and Clinical Outcomes Following Implantation?

The use of acellular dermal matrices (ADMs) in breast reconstruction is a controversial topic. Recent literature has investigated the effects of ADM sterilization on infectious complications, although with varying conclusions. Previous work by our group showed no difference between aseptic and sterilized products immediately out of the package. In this study, we investigate the microbiologic profiles of these agents after implantation. METHODS: In this prospective study, we cultured samples of ADM previously implanted during the first stage of tissue expander-based immediate breast reconstruction. A 1 cm2 sample was excised during the stage II expander-implant exchange procedure, and samples were incubated for 48 hours in tryptic soy broth. Samples with growth were further cultured on tryptic soy broth and blood agar plates. Patient records were also analyzed, to determine if ADM sterilization and microbial growth were correlated with infectious complications. RESULTS: In total, 51 samples of ADM were collected from 32 patients. Six samples were from aseptic ADM (AlloDerm), 27 samples were from ADM sterilized to 10-3 (AlloDerm Ready-to-Use), and 18 samples were from products sterilized to 10-6 (AlloMax). No samples demonstrated bacterial growth. Only 5 patients experienced postoperative complications, of whom only 1 patient was infectious in nature. We failed to demonstrate a statistically significant correlation between sterility and postoperative complications. CONCLUSIONS: Our findings showed no difference in microbial presence and clinical outcomes when comparing ADM sterility. Furthermore, no samples demonstrated growth in culture. Our study brings into question the necessity for terminal sterilization in these products.

The Effect of Axillary Lymph Node Sampling during Mastectomy on Immediate Alloplastic Breast Reconstruction Complications.

BACKGROUND: Tissue expander-based immediate breast reconstruction is currently the most common technique used for postmastectomy breast reconstruction. During mastectomy, axillary lymph nodes are biopsied to stage patients. The purpose of this study is to investigate postoperative complications with respect to extent of lymph node dissection. METHODS: A retrospective review of all patients undergoing tissue expander-based immediate breast reconstruction at our institution from 2010 to 2012 was conducted. Charts were analyzed to determine the association between the absolute number of axillary lymph nodes removed and postreconstructive incidence of skin necrosis, cellulitis, seroma, and expander removal. Independent sample t test and linear regression were used to analyze data. RESULTS: In total, 282 patients with 467 reconstructions were included. Overall incidence of all postoperative complications per breast was 23.8%. Breasts in which a complication occurred had a mean of 6 nodes removed versus 4 nodes in uncomplicated breasts (P = 0.018). Complications were noted at a significantly higher rate in patients who underwent axillary lymph node dissection compared with sentinel lymph node biopsy (P = 0.008). Expander removal and seroma occurred more frequently in breasts that had a greater number of nodes removed (P = 0.006 and P = 0.015, respectively). Preoperative radiation resulted in higher incidence of cellulitis and skin necrosis. Postoperative radiation and chemotherapy did not adversely affect reconstruction. CONCLUSIONS: Axillary lymph node removal of >4 nodes confers a greater risk of postreconstructive seroma formation and tissue expander loss in patients undergoing immediate reconstruction following mastectomy. Axillary lymph node dissection has a higher incidence of breast reconstruction complications compared with sentinel lymph node biopsy. Therefore, we encourage plastic surgeons to consider degree of lymphadenectomy when discussing reconstructive options with patients, as this may significantly impact their reconstructive outcome.

Revisiting Nasal Reconstruction After Mohs Surgery: A Simplified Approach Based on the Liberal Application of Local Flaps.

INTRODUCTION: Nasal reconstruction after oncologic resection often constitutes a technically demanding task. The aim of this study was to investigate our experience with nasal reconstruction after Mohs micrographic surgery using the following 3 well-established, versatile, and reliable techniques: the bilobed flap, nasolabial flap, and forehead flap. METHODS: A single-surgeon retrospective chart review of all patients undergone a bilobed, nasolabial, or forehead flap for nasal reconstruction after Mohs micrographic surgery from 2003 to 2017 was conducted. Data collected for analysis included demographics and defect characteristics. Revisions and complications were analyzed. RESULTS: A total of 83 patients were included in the study; 52 patients underwent bilobed, 17 patients nasolabial, and 14 patients forehead flaps. All defects involved the middle and lower nasal zones. There was a significant difference in the defect size between the 3 groups (P < 0.001). The mean diameters were as follows: 1.37 cm for the bilobed, 2.22 cm for the nasolabial, and 3.46 cm for the forehead flap groups. Liberal utilization of the bilobed and nasolabial flaps achieved reconstruction of defects larger than typically described in the literature. Reconstruction of all defects, including support for successful restoration of mucosal lining and cartilage when indicated, was achieved. An algorithm dealing with defects larger than 0.5 cm in the middle and distal nasal zones was devised. CONCLUSIONS: In this study, the liberal application of the 3 flaps was described based on an individualized treatment approach. The algorithm provides a simplified approach for larger nasal defects in the middle and lower thirds.

Digital Image Speckle Correlation to Optimize Botulinum Toxin Type A Injection: A Prospective, Randomized, Crossover Trial.

BACKGROUND: Historically, physicians have relied on their subjective measures when determining the site and dosages for botulinum toxin type A injections. Digital image speckle correlation is a technology that tracks pore movement from rest to maximal exertion, allowing for the determination of the optimal sites of injection. In this prospective, randomized, crossover trial, the efficacy of using digital image speckle correlation was compared to physician assessment in choosing botulinum toxin type A injection sites. METHODS: Ten female patients were analyzed in this blinded crossover study. Subjects were randomized to either injections based on digital image speckle correlation analysis or injections based on the 2004 facial aesthetics consensus recommendations. All patients received 20 U of botulinum toxin type A in the glabellar region and were crossed over and reinjected after 6 months. Follow-up was completed with the Facial Line Outcomes 11-item survey and repeated imaging with digital image speckle correlation, to measure patient satisfaction and degree of paralysis, respectively. Statistical comparison was completed by means of matched sample t test. RESULTS: On average, the digital image speckle correlation analysis provided 4.8 injection sites, whereas the practitioner chose five injections sites. Patients receiving digital image-directed injections had higher rates of satisfaction on the Facial Line Outcomes instrument (p = 0.0003) and a larger degree of paralysis (p = 0.003). Furthermore, muscle function returned to normal later in patients injected with digital image speckle correlation (17.9 weeks versus 20 weeks; p = 0.03). CONCLUSIONS: This study demonstrates the benefits of using digital image speckle correlation in determining optimal botulinum toxin type A injection location. Digital analysis allows practitioners to better treat facial rhytides by eliminating subjective decisions regarding dose and site of injection. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, II.

Comparison of the Basal View and a Previously Standardized Cleft Lip Rating Scale.

OBJECTIVES: Numerous scales assessing the aesthetics of cleft lip repair exist. Most, including the Asher-McDade scale, use frontal and lateral views, while neglecting a basal view. We believe this view is important for properly assessing the aesthetics of repair. In this study, we evaluate the basal view in comparison to the Asher-McDade scale. METHODS: This scale was based on progressive columellar shortening and alar flaring/slumping. A panel of 4 plastic surgeons applied the basal and Asher-McDade scales to pictures of patients with repaired unilateral cleft lip in 56 multiethnic participants aged 5 years to 18 years; images were scored from 1 to 5 (best). Statistical analysis was done via Spearman correlation. RESULTS: Scores from plastic surgeons correlated strongly for each view. There was moderate correlation for the basal view with both nasal form and deviation scores ( P < .05). As expected, there were no strong correlations between the basal view and vermillion border or profile. DISCUSSION: The Asher-McDade scale has been used to evaluate cleft lift repairs but is limited due to its subjective nature. The basal view scale grades these repairs by using a scale of progressive columellar shortening and alar flaring/slumping, which provides an opportunity for quantification and standardization. Our results show that the basal view correlates with the Asher-McDade scale among raters, thus providing an objective and validated measure of cleft lip repair.

Infectious Loss of Tissue Expanders in Breast Reconstruction: Are We Treating the Right Organisms?

BACKGROUND: Postoperative infections following tissue expander-based breast reconstruction represent a significant threat to the reconstructive process. Studies have found the incidence to be as high as 29%. There has been abundant research into the risk factors associated with these infections, although very few studies have focused on the causative organisms. The purpose of this study was to investigate the bacterial flora associated with tissue expander infections after breast reconstruction. METHODS: A retrospective analysis of all patients who underwent tissue expander-based breast reconstruction at our institution between February 2010 and April 2013 was conducted. The medical records were reviewed for demographic information, medical history, operative technique, postoperative course, and culture results. Descriptive data analysis was then performed. RESULTS: A total of 56 tissue expander infections were identified in 49 patients during the study period. 41.1% of the cultures grew gram-positive organisms, whereas 28.6% grew gram-negative species. The 2 most common organisms were Staphylococcus aureus (17.9%) and Staphylococcus epidermidis (14.3%). Pseudomonas aeruginosa was the most frequent gram-negative species and was also the third most frequent organism cultured (10.9%). DISCUSSION: Due to the high rate of infection in breast reconstruction patients, adequate perioperative prophylaxis is necessary. The surgeon must also be prepared to treat patients who may return with infection postoperatively. Although the majority of our infections were secondary to normal skin flora, a significant proportion were caused by gram-negative species. Given these results, the empiric antibiotic of choice for postoperative infections should be reconsidered to cover for these various organisms.

Is Sterile Better Than Aseptic? Comparing the Microbiology of Acellular Dermal Matrices.

INTRODUCTION: Postoperative infections are a major complication associated with tissue-expander-based breast reconstruction. The use of acellular dermal matrix (ADM) in this surgery has been identified as a potential reservoir of infection, prompting the development of sterile ADM. Although aseptic and sterile ADMs have been investigated, no study has focused on the occurrence and clinical outcome of bacterial colonization before implantation. METHODS: Samples of aseptic AlloDerm, sterile Ready-To-Use AlloDerm, and AlloMax were taken before implantation. These samples were incubated in Tryptic soy broth overnight before being streaked on Trypticase soy agar, MacConkey agar, and 5% blood agar plates for culture and incubated for 48 hours. Culture results were cross-referenced with patient outcomes for 1 year postoperatively. RESULTS: A total of 92 samples of ADM were collected from 63 patients. There were 15 cases of postoperative surgical site infection (16.3%). Only 1 sample of ADM (AlloMax) showed growth of Escherichia coli, which was likely a result of contamination. That patient did not develop any infectious sequelae. Patient outcomes showed no difference in the incidence of seroma or infection between sterile and aseptic ADMs. CONCLUSIONS: This study evaluates the microbiology of acellular dermal matrices before use in breast reconstruction. No difference was found in the preoperative bacterial load of either aseptic or sterile ADM. No significant difference was noted in infection or seroma formation. Given these results, we believe aseptic processing used on ADMs is equivalent to sterile processing in our patient cohort in terms of clinical infection and seroma occurrence postoperatively.