Cuticle Removal Research Articles

Decellularization: Leveraging a Tissue Engineering Technique for Food Production.

The edible nature of many plants makes leaves particularly useful as scaffolds for the development of cultured meat, where animal tissue is grown in the laboratory setting. Recently, we demonstrated that decellularized spinach leaves can serve as scaffolds to grow and differentiate cells for cultured meat products. However, conventional decellularization methods use solutions that are not considered safe for use in food, such as organic solvents (hexanes) and detergents (triton X-100 (TX100)). This study modified decellularization protocols to incorporate detergents that are regulated (REG) by the United States Food and Drug Administration (FDA) for use in food, such as Polysorbate 20 (PS20), and eliminates the use of hexanes for cuticle removal. Spinach leaves were decellularized with sodium dodecyl sulfate and then with either TX100 (control) or PS20. The average DNA content for TX100 samples and PS20 samples was similar (1.3 ± 0.07 vs 1.3 ± 0.05 ng/mg; TX100 vs PS20, p = ns). The importance of cuticle removal was tested by removing hexanes from the protocol. Groups that included the cuticle removal step exhibited an average reduction in DNA content of approximately 91.7%, and groups that omitted the cuticle removal step exhibited an average reduction of approximately 90.3% (p = ns), suggesting that the omission of the cuticle removal step did not impede decellularization. Lastly, primary bovine satellite cells (PBSCs) were cultured for 7 days (d) on the surface of spinach leaves decellularized using the REG protocol. After the 7 d incubation period, PBSCs grown on the surface of REG scaffolds had an average viability of approximately 97.4%. These observations suggest that the REG protocol described in this study is an effective decellularization method, more closely adhering to food safety guidelines, that could be implemented in lab grown meat and alternative protein products.

2023 TERMIS – AMERICAS Conference & Exhibition Boston Marriott Copley Place April 11–14, 2023

2023 TERMIS – AMERICAS Conference & Exhibition Boston Marriott Copley Place April 11–14, 2023

Acetic acid, vinegar, and citric acid as washing materials for cuticle removal to improve hatching performance of quail eggs

The cuticle is the outmost layer of the eggshell and may affect the hatchability by modulating eggshell conductance. Three different solutions using acetic acid (AA), vinegar (V), and citric acid (CA) for cuticle removal by egg washing were developed, and the effects of cuticle removal on hatching performance of quail hatching eggs were evaluated. A total of 5,238 fresh quail hatching eggs were randomly divided into 9 treatments as follows: unwashed control, nondipped (CND); washed control, water dipped (CWD); standard control, 0.13% sodium hyperchlorite (CSH); 2% AA (AA2); 4% AA (AA4); 44.4% V (V2); 88.8% V (V4); 2% CA (CA2); and 4% CA (CA4). Overall, AA4, V4, and CA4 treatments significantly improved the hatchability of fertile eggs (95.42%, 94.16% and 95.66%, respectively) (P < 0.05) and the hatchability of CND, CWD, CSH, AA2, V2 and CA2 treatments were 90.98%, 93.00%, 92.27%, 79.44%, 90.37%, and 90.59%, respectively. The eggshell thickness and cuticle quality results showed that all AA, V, and CA solutions can effectively remove the quail eggshell cuticle, and AA4, V4 and CA4 significantly decreased eggshell thickness (P < 0.05). Microbial activity on the eggshell surface in all acid treatments was reduced significantly at day 0 of incubation (P < 0.05) and that significantly decreased than controlled treatments over the incubation period except AA2 treatment.Egg weight loss was lower for all acid treatments than that of the CND treatment (P < 0.05). There was no clear effect of treatments on chick quality. Hatch time in AA4, V4, and CA4 treatments slightly improved compared with controlled treatments (P > 0.05). There were no significant differences between treatments for chick livability and live weight at the first 21 D of life. Results of the present study indicate that cuticle removal with AA4, V4, or CA4 could effectively decrease the microbial activity on the eggshell surface during the incubation period and improve hatchability of quail hatching eggs without negative effects on hatch time and performance of quail chicks.

Understanding the dynamics of keratin weakening and hydrolysis by proteases.

Keratin is the structural protein in hair, nails, feathers and horns. Keratin is recalcitrant, highly disulfide bonded and is generally inaccessible to common proteases. Only certain types of proteases, called keratinases, are able to cleave the peptide bonds within the keratin structure. Due to this outstanding activity, keratinases have potential application in industries such as livestock, cosmetics and pharmaceuticals. Yet, the process of enzymatic keratin degradation is poorly understood, affecting the development of industrial enzyme formulations that may require full or only partial modification or weakening. Here we investigate the dynamics of keratin weakening and hydrolysis, showing that the decrease in hair mechanical strength is associated with cuticle removal and damage to the cortex and complete breakdown is dependent on reducing agents. Proteases with keratinolytic activity were selected and applied to hair with degradation examined by mechanical, biochemical and microscopic techniques. The extent of keratin degradation was highly enhanced by the presence of reducing agents, principally sodium thioglycolate, exceeding 90% degradation within 16 h of enzymatic treatment. Application was extended to feathers showing that the findings are relevant to improving the use of keratinases in a variety of industries. Overall, the outcomes provide valuable insights into the keratin degradation process by enzymes for the optimization of cosmetic and pharmaceutical products and for livestock waste recycling among other important applications.

Rapid and effective cuticle removal from wool fibers using ionic liquid

Wool fibers are composed of cuticle and cortex cells, which are of obvious differences in many properties. The development of methods to isolate the two kinds of cells can provide platform to elucidate the roles they play in the characteristics of wool fibers. Here we demonstrate the cuticle can be completely and rapidly removed from the wool fibers by the use of ionic liquids, with inner cortex intact. Confocal microscope, SEM and FTIR have been applied to study the wool fibers after cuticle removal. In contrast to the traditional/long physical or chemical separation routes (>14 h), our method is very rapid (<1 h). This work demonstrates the ability of ionic liquid as a novel, rapid and efficient media for cuticle/ cortex isolation.

Comparison of microscopy techniques to visualize powdery mildew (Erysiphales) conidiophores.

Fast techniques allowing a clear view of fungal structures, specially conidiophores, are important for accurate diagnosis and identification at the genus level of powdery mildew fungi. In this study we propose a new technique for removing cuticles of leaves with a colorless nail base (cosmetic-based cellulose acetate) and transparent adhesive tape for viewing conidiophores of powdery mildew on papaya ( Carica papaya ), papayuelo ( Vasconcellea goudoutiana ), and yellow ipe ( Tabebuia serratifolia ) leaves by light microscopy. For comparison, the traditional technique of discoloration (diaphanization) of the leaves was also tested. Phase and interference contrast microscopy and liquid montant with stains were used to improve the optical contrast. The cuticle removal technique showed to be simple and easy to use, showing more clearly the conidiophores origin and the leaf topology thus evidencing that it can be used as an auxiliary routine technique for the identification of powdery mildew fungi.

Prevalence of lymphedema in women undergoing treatment for breast cancer in a referral center in southeastern Brazil

BackgroundLymphedema is a highly prevalent condition in women who have undergone treatment for breast cancer. Lymphedema negatively affects the quality of life. The objective of this study was to estimate the prevalence of lymphedema and associated factors in women treated for breast cancer in the municipality of Juiz de Fora.MethodsWe performed a cross-sectional study that evaluated 250 women who were being treated for breast cancer. Pre-screening of the sample by analysis of medical records was performed to select women who met the inclusion criteria as follows: women who had an operation more than 6 months ago; absence of active disease, locoregional or distant; the absence of functional change in the affected limb before surgery, which could lead to swelling of the limb; and simulating or masking symptoms of lymphedema, such as bursitis, tendonitis, and work-related musculoskeletal disorders. Women with bilateral breast cancer, absence of axillary intervention (partial or complete axillary dissection and/or SLN biopsy), active disease in the region, or lympho-venous alteration of the limb before surgery were excluded. Data were collected from the medical records of the selected cases, and they subsequently underwent an interview and a physical assessment.ResultsThe prevalence of lymphedema was 44.8%. There were medical records on the presence of this condition in 5.4% of cases. With regard to shoulder joint mobility, restrictions on abduction movements, internal and external rotation, and anterior shoulder adduction were significantly associated with lymphedema. Variables, including the presence of seroma, vascular changes, time elapsed after surgery, episodes of redness in the extremities, and cuticle removal from the hand with pliers were considered as major associated factors for lymphedema (p<0.05).ConclusionsThe prevalence of 44.8% for lymphedema found in this study is considered to be relevant because it is a morbidity that produces psychological, physical, and functional damage in patients with this condition. The planning of health programs and services appropriate for the immediate postoperative treatment of women with breast cancer, and increasing the awareness of health professionals regarding the early diagnosis of lymphedema, can help minimize the morbidity of this disease.

Line-scan hyperspectral imaging for real-time in-line poultry fecal detection

The ARS multispectral imaging system with three-band common aperture camera was able to inspect fecal contaminants in real-time mode during poultry pro- cessing. Recent study has demonstrated several image processing methods including binning, cuticle removal filter, median filter, and morphological analysis in real-time mode could remove most false positive errors. The ARS research groups (Beltsville, MD and Athens, GA) and their industry partner are now merging the fecal detection and systemically disease detection systems onto a common platform using a line-scan hyperspectral imaging system. This system will aid in commercialization by creating one hyperspectral imaging system with user-defined wavelengths that can be installed at different locations of the processing line to solve significant food safety prob- lems of disease as well as contaminant detection. Thus, this research demonstrated the feasibility of line-scan hyper- spectral imaging system in terms of processing speed and detection accuracy for a real-time, in-line fecal detection at current processing speed (at least 140 birds per min) of commercial poultry plant. Our preliminary results showed the real-time hyperspectral imaging system could detect small amount (about 10 mg) of fecal (duodenum, cecum, colon) and ingesta contaminants. The system performance could be improved by optimizing line lighting system especially near-infrared bands for quality images and additional spectral images to minimize false positive detection errors. The newly developed line-scan hyper- spectral imaging system could improve Food Safety Inspection Service (FSIS)'s poultry safety inspection pro- gram significantly.

Alternative Method for Hazelnuts Peeling

The paper proposes an improved peeling treatment for hazelnut fruits consisting in dipping them in a solution (NaOH 0.4 g/100 g, NaH2PO4·H2O 0.2 g/100 g) for 10 min at 50 °C, then scrubbing them with rubber rollers. The effects of the treatment on quality and peeling of four Italian hazelnut varieties were investigated. The data collected on peeling capability showed that the treatment proposed was effective in cuticle removal with a yield of peeling higher than 90%. The comparison of chemical–physical parameters in unpeeled and peeled hazelnut samples highlights that no difference was detected in texture, sugar, protein and α-tocopherol values. After the peeling process a significant change (p < 0.05) in colour, total fat and peroxide value was observed in peeled hazelnut samples when compared with the unpeeled ones. With regard to the sensorial parameters, no significant changes were found between unpeeled and peeled hazelnut samples. The peeling solution proposed in this study shows a good potential in industrial applications as an alternative to the usual manufacturing peeling method (roasting) that causes changes, not always desired, to some sensorial and chemical–physical parameters of the fresh fruit.

In-plant Test of In-line Multispectral Imaging System For Fecal Detection during Poultry Processing

Food Safety Inspection Service established new requirements for the zero tolerance for fecal contaminated broiler carcasses entering the chiller. The hyperspectral imaging research accomplished at ARS has demonstrated the real-time multispectral imaging system was able to effectively detect fecal contaminants during poultry processing. The imaging system with two-band (517 and 565 nm) was able to inspect every bird in real-time, in-line mode at commercial processing speed. With several real-time image processing methods including binning, cuticle removal filter, median filter, and morphological analysis, the system performance was improved by maximizing detection accuracy and minimizing false positive errors. Most false positives were found at wing bone, thigh, vent, and boundary. Since the number of true positives confirmed organoleptically by human inspector was only 14 out of 29,821 carcasses (0.05%), more samples need to be tested to confirm the performance of detection accuracy. Overall, the ARS multispectral imaging system was able to detect contaminants with high detection accuracy (approximately 91%) and 3.3% false positive errors at the processing speed of 150 birds/min from the two-day in-plant trials. However, the performance of ARS real-time in-line imaging system for fecal contaminant detection on broiler carcasses can be improved by adopting a better camera which has higher signal-to-noise ratio such as line-scan real-time hyperspectral imaging system currently being investigated for its applicability for real-time processing.

Adaptive Image Processing Methods for Improving Contaminant Detection Accuracy on Poultry Carcasses

A real-time multispectral imaging system has demonstrated a science-based tool for fecal and ingesta contaminant detection during poultry processing. In order to implement this imaging system in the commercial poultry processing industry, the false positives must be removed. To do this, we tested and implemented additional image processing algorithms including binning, cuticle removal filter, median filtering, and morphological analysis in real-time mode to maximize detection accuracy and minimize false positives (FPs). The median filtering and binning processes were able to reduce FPs up to 98.7% and 95.2%, respectively, by eliminating most of the salt-and-pepper noise from the raw images. The detection accuracy varied with the parameter values of the image processing algorithms, including binning, thresholding, median filtering, and morphological analysis. Overall contaminant detection accuracy on moving birds varied from 84.3% to 97.8%, and FP errors were 1.9% and 41.8%, respectively. Although neither the overall detection accuracy nor FP errors were affected by the camera gain, the detection accuracy results changed slightly, from 87.4% to 95.1%. In this case, the FP errors were 1.8% and 15.9%, respectively. Thus, the USDA-ARS multispectral imaging system was able to detect contaminants with 91.6% accuracy and 3.3% FP errors by selecting optimum image processing methods at a processing speed of 140 birds per minute.

Human hair: color changes caused by daily care damages on ultra-structure

The relation between hair ultra-structure damages and color changes was studied. Virgin dark-brown hair was hand-washed, using lauryl sodium sulfate solution in 40 °C water, rinsed, wet-combed, heat-dried and dry-combed for up to 120 times. Ultra-structure changes were studied by electron microscopy. The treatments damage the cuticle and the cortex. The extraction of soluble material renders cavities, or holes, in the endocuticle. The cavities are 50–200 nm in diameter. There are two kinds of cavities: some filled with lower density material than the remaining endocuticle and some filled with air or water vapor. Displacement, cracking and cleavage of cuticle cells are also observed. Cuticle removal was found to proceed in two ways: via cleavage through the cell membrane complex, and via endocuticle rupture, taking place preferentially in the cavities’ surroundings. In the cortex, cavities develop in the intermacrofibrilar cement, in the cell membrane complex and around the melanin granules. These ultra-structural damages give rise to significant changes on hair color, as shown by diffuse reflectance spectrophotometry. The hair lightness was found to increase after soft washing treatments (5–20 washes), or after keeping it in 40 °C water. Deeper hair degradation turns the hair lightness undistinguishable from the initial value, but changes the color mainly by a yellowing of the hair. A simple model based on light reflection was developed to explain hair reflectance behavior before and after damage; results show a reasonable agreement with the experimental data.

Response of In Situ Leaf Psychrometer to Cuticle Removal by Abrasion

In situ measurements of leaf water potential by thermocouple psychrometers are useful in monitoring the energy state of water in plants. Excessive equilibration times caused by low vapor conductance across the leaf cuticle and vapor sorption in the psychrometer chamber often make in situ sampling difficult. Several methods of removing the waxy leaf cuticle have been shown to increase water vapor conductance across the epidermis of the leaf, but their relative effectiveness has not been compared and equilibration times are unknown. Six wax removal treatments were compared with an untreated control to determine the most effective technique for increasing water vapor conductance across the leaf surface. The mean time for the leaf psychrometer to reach an equilibrated water potential was estimated for the treatment that produced the greatest conductance. Abrasion treatment of leaves with 600 grit sandpaper produced the greatest increase in cuticular conductance, resulting in a mean equilibration time for psychrometer measurements of 7.5 min.

Effects of Breeder Dietary Fat and Eggshell Cuticle Removal on Subsequent Broiler Growout Performance

Broiler breeder hens were fed isocaloric diets containing one of three different types and levels of added fat. Eggs collected when hens were 52 and 62 wk of age were either left intact or were washed to remove the eggshell cuticle prior to set. Chicks hatched from each breeder diet and eggshell cuticle treatment were reared through 22 days to assess effects on broiler performance. An increase in feed consumption in broilers from 52-wk-old breeders was associated with cuticle removal. It was concluded that eggshell cuticle removal in eggs from hens during post-peak production may lead to subsequent increases in feed consumption of broiler offspring.

Effects of dietary fat and eggshell cuticle removal on egg water loss and embryo growth in broiler hatching eggs

The effects of dietary fat and eggshell cuticle removal on egg water loss, embryo growth, and hatchability were determined in eggs from broiler breeder hens at different ages. Hens were fed isocaloric diets containing one of three different types and levels of added fat. In addition, eggs were either left intact or washed to remove the eggshell cuticle prior to set. Cuticle removal increased egg water loss between 43 and 62 wk. Cuticle removal increased relative wet embryo weight at Week 52 and relative dry embryo weight at 52 and 62 wk. Furthermore, at 62 wk, diet and day of incubation interacted to affect wet embryo weight, and diet variably affected dry embryo weight. No treatment differences were observed for cumulative hatchability, rate of hatch, and relative yolk sac weight at Day 19 of incubation. It was concluded that cuticle removal and the addition of fat to breeder diets may influence embryonic growth without having any subsequent effects on hatchability.

Automated Spray Sanitizing of Broiler Hatching Eggs 1. Physical Characteristics of The Egg

This study examines the physical effects of spray sanitizing nest clean broiler hatching eggs with the MS Technologies, Inc. Mini-Master/4000® or by immersion dipping immediately following collection. The measurements taken during sanitizing were air, shell surface, and yolk surface temperatures. Researchers studied eggshell conductance, percentage egg moisture weight loss, and cuticle integrity following sanitizing. Treatments included non-sanitized controls and eggs sanitized by water dipping or automated spray washed with water, or spray washed and sanitized with chemical solutions. The automated spray washing consisted of two stages. In Stage 1, the temperature of the washing solution was 111°F, and in Stage 2 the temperature of the sanitizing solutions was 118°F. Automated spray sanitizing at fast processing speed (16 sec per flat) caused air and shell surface temperatures to rise by 28.4° and 20.3°F, respectively. These temperatures returned rapidly to pre-treatment temperature (83°F). Dipping eggs for 2 min in a sanitizer solution at 111°F resulted in a pattern of air and shell surface temperature increase and recovery similar to eggs sanitized by automation. Automated spray sanitizing caused a reduction in the shell cuticle, but cuticle removal was not severe enough to significantly increase eggshell conductance or percentage egg moisture weight loss.

Effect of Altering Eggshell Permeability on the Hatchability of Turkey Eggs Incubated at High Altitude ,

Factors of Fick's First Law of Diffusion were manipulated in order to observe their effect on the hatchability of turkey eggs at high altitude. Shell permeability was studied by removing the eggshell cuticle. The eggshell surface for gas exchange was studied at different times of the laying cycle as the surface area increased. Differences in gas tension across the shell were assessed under two oxygen pressures (131 and 109 ton of oxygen).Increasing shell permeability in order to increase gas flow to the embryo did not improve hatchability but did increase (P≤.05) embryonic mortality. On Day 25 these data suggest that cuticle removal is not a good means for improving the hatchability of turkey eggs at high altitude. Eggs from older hens had increased surface area and decreased hatchability at high altitude.An oxygen partial pressure of 109 torr depressed (P<.05) hatchability, compared to that resulting from a partial pressure of 131 torr. The depression in hatchability was attributed mainly to embryos that were unable to free themselves from the shell once they had pipped.

TEM and X-Ray Photoelectron Spectroscopic Studies of Wool Fibers after Cuticle Removal

A method is described for cleanly separating the cortex and cuticle of wool fibers using water or an aqueous buffered solution as the immersion medium. Nearly complete removal of cuticle cells can be achieved in 2–3 hours when snippets of wool are vortex-mixed with a suspension containing both glass fiber snippets and glass beads. Examination of treated fibers (transverse sections) by transmission electron microscopy indicates removal of cuticle cell layers (endo- and exocuticle), together with associated membrane intracellular bands ( i-layers). The surface elemental composition of the fiber (determined using x-ray photoelectron spectroscopy) changes appreciably with cuticle removal. The sulfur content of the intracellular cortical proteins at the exposed surface is estimated to be 2%, compared to 9% for the epicuticle proteins at the surface of untreated wool.

Effect of cuticle removal on the water vapour conductance of egg shells of several species of domestic bird

Abstract 1. The effect of egg shell cuticle removal, using a 14.7 mM solution of sodium hypochlorite, on the water vapour conductance of egg shells was studied in 5 species of domestic bird. 2. The water vapour conductance of fowl and Muscovy duck egg shells was not affected by the treatment. 3. Cuticle removal increased the conductance of turkey and goose shells, particularly those already relatively porous to water vapour. 4. Removal of cuticles from domestic duck egg shells increased water vapour conductance. In this case the effect was most marked for eggs with lower initial conductances. 5. The differing patterns of resistance to water vapour loss associated with egg shell cuticle are related to the ultrastructure of the cuticle itself, especially the number and size of cuticular fissures. 6. The results suggest that egg shell cuticle is a component in a series of resistances associated with the egg shell and that the use of Fick's Law is insufficient to explain water vapour diffusion across the shel...

Effects of Eggshell Cuticle Removal and Incubation Humidity on Embryonic Development and Hatchability of Broilers

The effects of eggshell cuticle removal and two levels of incubation humidity 28.3 C [50% relative humidity (RH)] and 30.0 C (55% RH) wetbulb temperature (WB) on embryonic mortality and hatchability were determined from broiler hatching eggs laid during 38, 42, 48, and 54 weeks of age. Variables measured were: egg weight loss during the first 17 days of incubation, hatch at Days 19.5 and 20.5 of incubation, hatch of fertile eggs, stage of embryonic mortality, and chick weight at 21.5 days of incubation.Day 0 to 17 percentage egg weight loss was increased when the incubation humidity was lowered and the loss was greater than that observed after cuticle removal. A greater percentage of chicks hatched on Day 19.5 at 28.3 C than at 30.0 C WB. The percentage hatch of 38-week fertile eggs was improved at the higher humidity; the higher humidity also decreased late dead and increased pipped embryonic mortalities. Cuticle removal decreased early dead and increased late dead mortality. At Week 38 cuticle removal and lower humidity resulted in a decrease in chick weight at 21.5 days of incubation. For Weeks 42, 48, and 54 combined, pipped mortality was increased by higher humidity and late dead mortality was increased by cuticle removal.Water loss from the egg was increased by cuticle removal or by lowering incubation humidity from 30.0 C to 28.3 C WB, or by both, but lowering humidity was more effective. Changes in humidity and cuticle removal may affect vital gas exchange to different degrees; however, the effect of cuticle removal on embryonic development may be dependent upon the quantity and morphology of the cuticle. Cuticle removal may provide a means to improve vital gas exchange if appropriate adjustments in humidity are made to prevent excessive water loss.