Search

Objective: To examine trends, characteristics, and outcomes of donor oocyte embryo transfer cycles by original oocyte and resultant embryo state and determine whether oocyte state (fresh or frozen) is differentially associated with clinical pregnancy, live birth, and term, healthy birthweight neonates among singleton live births., Design: Retrospective cohort study., Setting: National study., Patient(s): Patients undergoing donor oocyte embryo transfer cycles in the United States reporting to the National Assisted Reproductive Technology Surveillance System from 2013 to 2020 . INTERVENTION(S): Original donor oocyte and resultant embryo state (fresh or frozen)., Main Outcome Measure(s): Annual numbers and proportions of total donor oocyte embryo transfer cycles stratified by oocyte and embryo state and single embryo transfer cycles resulting in the live birth of term (≥37 weeks gestation), healthy birthweight (≥2,500 g) singletons during 2013-2020. Rates of live birth and term, healthy birthweight neonates among singleton live births for 2018-2020 are also reported. Relative risks examine associations between donor oocyte state and live birth and term, healthy birthweight neonates among singleton live births resulting from donor oocyte embryo transfer cycles., Result(s): From 2013 to 2020, there were 135,085 donor oocyte embryo transfer cycles, of which the proportions increased for frozen embryos (42.3%-76.6%), fresh embryos using frozen donor oocytes (19.9%-68.3%) and single embryo transfers (36.4%-85.5%). During 2018-2020, there were 48,679 donor oocyte embryo transfer cycles. Rates of live birth were lower with frozen compared with fresh donor oocytes for both fresh (46.2%, 55.9%; adjusted relative risk [aRR], 0.83; 95% confidence interval [CI], 0.79-0.87) and frozen (41.3%, 45.8%; aRR, 0.94; 95% CI, 0.91-0.98) embryo transfer cycles. Among singleton live births, rates of delivering a term, healthy birth weight neonate were similar for frozen compared with fresh donor oocyte transfer cycles among fresh (77.3%, 77.2%; aRR, 1.01; 95% CI, 0.98-1.03) and frozen (75.6%, 75.1%; aRR, 1.02; 95% CI, 0.99-1.04) embryos., Conclusion(s): In this national study of donor oocyte embryo transfer cycles, frozen embryo transfers, fresh embryo transfers using frozen oocytes, and single embryo transfers increased. Although frozen compared with fresh oocytes were associated with a slightly reduced rate of live birth, rates of term, healthy birthweight neonates among singleton live births were comparable between donor oocyte states., Competing Interests: Declaration of Interests C.B.B. reports funding from Marianne Ruby, MD Award from the Department of Gynecology and Obstetrics, Emory University School of Medicine for a senior medical student to conduct research in an area that will advance knowledge and understanding of issues impacting women from menarche through menopause; and the Pacific Coast Reproductive Society Scholarship for part of this research to be presented at the Annual Pacific Coast Reproductive Society Conference in Indian Wells, California, March 22–26, 2023. C.E.D. has nothing to disclose. J.C.L. has nothing to disclose. D.M.K. has nothing to disclose. J.F.K. has nothing to disclose., (Copyright © 2024 American Society for Reproductive Medicine. All rights reserved.)

Lymphoproliferative disorders in pregnancy are rare. Among that, Castleman disease is an uncommon lymphoproliferative disorder in pregnancy with an unknown etiology. Many cases have not been encountered more often in practice and are easily misdiagnosed due to their rarity. We present a case of a 36-year-old pregnant woman with a left para-ovarian mass incidentally diagnosed during the elective caesarean section as a benign lymphoproliferative disorder, sent in frozen. Diagnosing Castleman disease in frozen histopathology is quite challenging for pathologists, and few cases have been reported., Competing Interests: Human subjects: Consent was obtained or waived by all participants in this study. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Ramamoorthy et al.)

In order to explore the effect of sub-freezing storage on water holding capacity and tenderness of beef, four treatments were compared in this study: sub-freezing (-7 °C) fast sub-freezing (-38 °C until the core temperature achieved to -7 °C), superchilling (-1 °C) and fast frozen (-38 °C until the core temperature achieved to -18 °C) with the latter two treatments serving as the controls. The differences in muscle fiber structure, water distribution, protein oxidation and cytoskeletal protein degradation were studied. The results demonstrated that compared with other treatments, the fast sub-freezing treatment resulted in less structural damage to the muscle fibers and had better water holding capacity. Both sub-freezing and fast sub-freezing treatments inhibited protein oxidation compared with superchilling, but the former treatment's level of protein oxidation was higher than that in fast sub-freezing treatment during long-term storage (42 weeks). In addition, the structural proteins in the sub-freezing and fast sub-freezing treatments underwent faster degradation during long-term storage and therefore the meat was more tender compared with the fast frozen treatment. The results indicate that the fast sub-freezing treatment can be potentially applied in beef storage., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study aimed to investigate near-infrared spectroscopy (NIRS) in combination with classification methods for the discrimination of fresh and once- or twice-freeze-thawed fish. An experiment was carried out with common carp ( Cyprinus carpio ). From each fish, test pieces were cut from the dorsal and ventral regions and measured from the skin side as fresh, after single freezing at minus 18 °C for 15 ÷ 28 days and 15 ÷ 21 days for the second freezing after the freeze-thawing cycle. NIRS measurements were performed via a NIRQuest 512 spectrometer at the region of 900-1700 nm in Reflection mode. The Pirouette 4.5 software was used for data processing. SIMCA and PLS-DA models were developed for classification, and their performance was estimated using the F1 score and total accuracy. The predictive power of each model was evaluated for fish samples in the fresh, single-freezing, and second-freezing classes. Additionally, aquagrams were calculated. Differences in the spectra between fresh and frozen samples were observed. They might be assigned mainly to the O-H and N-H bands. The aquagrams confirmed changes in water organization in the fish samples due to freezing-thawing. The total accuracy of the SIMCA models for the dorsal samples was 98.23% for the calibration set and 90.55% for the validation set. For the ventral samples, respective values were 99.28 and 79.70%. Similar accuracy was found for the PLS-PA models. The NIR spectroscopy and tested classification methods have a potential for nondestructively discriminating fresh from frozen-thawed fish in as methods to protect against fish meat food fraud.

Red blood cell (RBC) transfusion is a critical therapy for those with sickle cell disease (SCD). Alloimmunization is frequent for those with SCD and may limit the availability of matched RBC. Cryopreserved RBCs, from family members or donors with a similar RBC antigen profile could provide a viable alternative to avoid further alloimmunization and prevent hemolytic transfusion-related events. However, cryopreserved SCD and Sickle Cell trait (S-trait) donor RBC units suffer from reduced recovery following deglycerolization. This study proposes and tests a modified deglycerolization protocol using an automated cell processor to mitigate RBC loss. Six red cell concentrates (RCC) from donors with S-trait and six control RCCs were glycerolized, frozen (<-65 °C) and deglycerolized on the ACP 215 using modified parameters (decreased hypertonic solution flow rate (100 mL/min) and hypertonic equilibration delay (120 s), and increased NaCl dilution volumes (500 mL). Quality testing included: hematocrit (HCT), hemolysis, indices, extracellular potassium, morphology, osmotic fragility, osmotic gradient ektacytometry, hemoglobin (HGB), and recovery. Canadian standards (CS) indicate that acceptable deglycerolized units for transfusion require a HCT ≤0.80 L/L, HGB ≥35 g/unit, and hemolysis <0.8 % in 90 % of units tested. No significant differences in HGB or RBC recovery were observed between study groups. Significant differences between study groups were identified in osmotic fragility and osmotic gradient ektacytometry parameters. Of the 6 S-trait RCCs, 3/6 units were within the HCT, HGB and hemolysis thresholds set by the CS. The modified deglycerolization protocol provides a path for the routine cryopreservation of S-trait RBCs., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Biological samples of lipids and metabolites degrade after extensive years in -80 °C storage. We aimed to determine if associated multivariate models are also impacted. Prior TOFI&#95;Asia metabolomics studies from our laboratory established multivariate models of metabolic risks associated with ethnic diversity. Therefore, to compare multivariate modelling degradation after years of -80 °C storage, we selected a subset of aged (≥5-years) plasma samples from the TOFI&#95;Asia study to re-analyze via untargeted LC-MS metabolomics. Samples from European Caucasian ( n = 28) and Asian Chinese ( n = 28) participants were evaluated for ethnic discrimination by partial least squares discriminative analysis (PLS-DA) of lipids and polar metabolites. Both showed a strong discernment between participants ethnicity by features, before (Initial) and after (Aged) 5-years of -80 °C storage. With receiver operator characteristic curves, sparse PLS-DA derived confusion matrix and prediction error rates, a considerable reduction in model integrity was apparent with the Aged polar metabolite model relative to Initial modelling. Ethnicity modelling with lipids maintained predictive integrity in Aged plasma samples, while equivalent polar metabolite models reduced in integrity. Our results indicate that researchers re-evaluating samples for multivariate modelling should consider time at -80 °C when producing predictive metrics from polar metabolites, more so than lipids., Competing Interests: Aidan Joblin-Mills, Zhanxuan E. Wu and Karl Fraser are/were employees of AgResearch Limited The paper reflects the views of the scientists and not the company.

Extending the shelf life of exported beef could increase international demand and producer profits. The objective was to evaluate the effects of topically applying combinations of acerola cherry powder and rosemary extract on the shelf life of frozen-thawed bone-in beef short rib and chuck roll steaks. Chuck rolls (IMPS 116A; N = 9) and bone-in short ribs (IMPS 123A; N = 18) were aged (7 d; 0 °C), frozen (30 d; -20 °C), and thawed (60-72 h; 0 °C). Steaks measuring 1.02 cm thick were treated and subjected to a 4 d retail display. Steaks were left untreated (control) or sprayed topically with acerola cherry powder (0.05%; A), rosemary extract (0.10%; R), or a combination (M1 = 0.05% A + 0.1% R; M2 = 0.1% A + 0.1% R; M3 = 0.05% A + 0.2% R; M4 = 0.1% A + 0.2% R). Chuck roll M2- and M4-treated steaks were redder than the control steaks on days 3 and 4 ( p = 0.008), and antioxidant-treated steaks had less lipid oxidation on day 4 than the control steaks ( p = 0.021). Bone marrow samples treated with R, M3, and M4 were redder than the control on days 1-3 ( p = 0.014), and bone marrow treated with M3 was subjectively redder compared to the control on days 0 and 1 ( p = 0.033). Topical antioxidants improve the redness and delay the oxidation of frozen-thawed beef.

Differences between the stability of α-, β-, γ-, and δ-tocopherol as well α-tocotrienol stored at -20 °C and -80 °C were studied in broccoli and blueberry samples. Before storage up to 28 days, they underwent different initializing processes such as freezing quickly with liquid nitrogen and freeze-drying, followed by homogenization. While α-tocopherol levels in blueberries did not significantly differ, levels in broccoli were substantially higher after homogenization of freeze-dried samples compared to fresh broccoli samples. This might be caused by higher extractability of α-tocopherol from the changed cell structure. Storage of fresh broccoli samples at -20 °C led to decreasing α-tocopherol levels. Nevertheless, the deviation between freeze-dried samples to the initial fresh samples and fresh samples frozen with liquid nitrogen stored at -20 °C for 7 days were in the same order of magnitude. In conclusion, storage up to 7 days for vitamin relevant samples before analysis seemed to be justifiable., Competing Interests: There has been no conflict of interest for any author., (© 2024 The Authors. Published by Elsevier Ltd.)

Premature lymphocytes develop into non-autoreactive, mature naïve CD4 + or CD8 + T cells in the thymus before entering the circulation. However, in-depth characterization of human thymocyte development remains challenging due to limited availability of human thymus samples and the fragile nature of thymocyte populations. Thymocytes often do not survive cryopreservation and thawing procedures, especially the fragile CD4 + CD8 + double positive population. It is generally recommended to use fresh human thymus tissue on the day of excision to avoid any biases in thymocyte composition. This hampers the possibility to perform multiple experiments on the same thymus sample. To establish how the thymocyte viability and composition can be maintained, we compared two thymocyte isolation methods used for human and/or mice thymi, three cryopreservation methods in combination with our most gentle thawing technique. Based on our findings we established that fresh human thymi remain viable in cold storage for up to two days post-surgery without compromising thymocyte composition. Thymocytes can be cryopreserved if required, although the CD4 + CD8 + double positive populations may be reduced. Our study provides thoroughly optimized methods to study human thymocyte development over a considerable time-frame post-surgery., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Objective: To compare Frozen Section (FS) results during the reimplantation stage of revision knee arthroplasty, in patients without clinical signs of infection but with preoperative inconclusive serum inflammatory markers., Methods: Sections were revisited the day after surgery. Intraoperative FS (iFS) was accepted as positive when the presence of >5 polymorphonuclear neutrophils (PMNLs) in 5 separate high-power fields was determined according to the consensus criteria of the International Consensus on Musculoskeletal Infection. The clinical outcomes, cultures and diagnostic values of iFS and review FS (rFS) were analyzed., Results: No complications developed after reimplantation in 66 (84.6%) of the 78 evaluated patients. Complications developed in 12 patients, six of whom were treated with re-explantation, four with arthrodesis and two with above-the-knee amputation. Both iFS and rFS yielded insignificant sensitivity and specificity (25% and 45.5%, 25% and 45%, respectively). There was no statistically significant difference between definitive culture and iFS and rFS., Conclusion: iFS evaluation is insufficient to exclude recovery from periprosthetic joint infection (PJI). Diagnosis of recurrence of infection in patients with indefinite serum inflammatory markers between the explantation and reimplantation interval remains challenging due to massive fibrosis that makes proper tissue sampling difficult. The attending physician should closely monitor clinical findings., Competing Interests: No conflict of interest was declared by the authors., (© Copyright 2024 by Istanbul Provincial Directorate of Health.)

Purpose: This study aims to investigate the effects of frozen storage on the stability of traditional dough starters in China., Methods: The microbial community structure and abundance of related metabolic genes in different fermented sourdough prepared by Jiaozi (JZ) and Laomian (LM) starters before and after frozen storage at -20°C for half a year were analyzed using the shotgun metagenomic sequencing method, and differences in characteristics of texture in steamed bread were also compared by formal methods., Results: The fermentation ability (FA) and metabolic activities of yeast in the JZH sourdough (started by JZ which was stored at -20°C for half a year) were better than those of LMH sourdough (started by LM which was stored at -20°C for half a year). The dominant genera of Acetobacter were found to be increased in the JZH0 sourdough (started by JZH and fermented for 0 h) and those of Lactobacillus were found to be decreased. Lactobacillus (98.72%), Pediococcus (0.37%), Saccharomyces (0.27%), and Acetobacter (0.01%), were dominant in sourdough LMH0 (started by LMH and fermented for 0 h). The abundances of "oxidative phosphorylation-related enzymes" and the "biosynthesis of glutamate"-related enzymes and genes related to "biosynthesis of glutamate" and "unsaturated fatty acid" were higher in JZH0 than in the JZ0 sourdough (started by JZ without being frozen and fermented for 0 h). The good FA of yeast, the acid production capacity of bacteria in the sourdough, and the quality of the JZH steamed bread (made by the JZH starter) indicated the better freezing tolerance of the microorganisms in JZ than in LM., Conclusion: The conclusion of this study suggests the better application potential of the JZ as the fermentation starter in actual production., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Li, Lv, Zhang, Wang, Yang and Qu.)

Atlantic mackerel ( Scomber scombrus ) caught during the summer months in Icelandic waters after intensive feeding is rich in lipids and, thus, sensitive to lipid degradation. Recent studies have led to improved cooling and handling on board, ensuring high-quality raw material. However, studies on the development of high-quality products for human consumption are lacking. The study aimed to investigate the effects of hot-smoking on the physicochemical, microbial, and sensory quality of deep-skinned Atlantic mackerel fillets during chilled storage (1 ± 0.6°C). In addition, the quality of smoked mackerel from frozen-thawed fillets (9 months at -25 ± 1.8°C) was compared to that of fresh-smoked fillets to evaluate the possibility of the industry being able to provide smoked fillets throughout the year, despite the short fishing season. Brining and hot-smoking reduced total viable counts and inactivated Listeria monocytogenes . Hot-smoking positively affected the sensory attributes of the fillets and sensory quality was largely maintained for at least 21 days of chilled storage. Although slightly lower sensory and texture scores were obtained for frozen-thawed smoked fillets, they remained within acceptable limits throughout the period of cold storage. The shelf-life of smoked Atlantic mackerel deep-skinned fillets stored at 1°C is, therefore, assessed to be at least 21 days. Well-fed Atlantic mackerel is suitable for developing high-quality and stable smoked fillet products from both fresh and frozen-thawed raw materials., Competing Interests: The authors declare that they do not have any conflict of interest., (© 2024 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

Mucormycosis usually occurs in immunocompromised patients or those with uncontrolled diabetes. Along the third wave of SARS-CoV-2, an associated angioinvasive opportunistic infection with Mucor, a life-threatening fungal infection, was rampant and emerging. With an increase in the usage of steroids in the COVID scenario, the rate of mucormycosis did take a rapid and alarming increase in King Edward Memorial Hospital, Pune, India. Any delay in the diagnosis and management of the disease was life-threatening. The most conventional methods to diagnose mucormycosis are microbiological culture and histopathology of the tissue. The microbiological culture method plays an important role in the diagnosis of mucormycosis. However, the technique is labour-intensive, taking seven to eight days. Histopathology leads to false-negative reports if the tissue is not biopsied from representative sites. On the other hand, molecular methods are rapid, reliable, and applicable to different body samples, such as tissue, paraffin-embedded tissue blocks, plasma, and urine. We aimed to use a reverse transcriptase polymerase chain reaction (RT-PCR) method to detect Mucor in plasma samples. Due to a lack of availability of fresh samples, nucleic acid was extracted from the tissue sections of 69 cases diagnosed as Mucor by histopathology. These samples were subjected to RT-PCR using the MucorGenius kit (Pathonostics, Maastricht, Netherlands). A total of 57 tissue samples were sent for culture, and 49% of our cases were positive by culture and equally by RT-PCR. There was 80% sensitivity and 76% specificity between culture and PCR techniques. However, the use of blood/plasma for RT-PCR for early diagnosis of mucormycosis will be the method of choice., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Kiruthiga et al.)

Aim and Objectives: To estimate and evaluate the optimal thickness of tissue section for skin and dermal appendages intra-operatively and to compare the morphology and architecture of the epidermis and its appendages at 7 and 10 microns., Methodology: After obtaining clearance from the institutional human ethical committee, 101 skin margins were prospectively analyzed using a two-step embedding technique. After multiple trials and errors, 7- and 10-micron thicknesses were selected for the present study. Artefacts, staining characteristics, cellular morphology, cellular outline, and nuclear outline were assessed and scored as unacceptable or acceptable. The data were entered in a Microsoft Excel spreadsheet and analyzed using SPSS software., Results: There was a statistically significant difference between the sections obtained at 7 microns and 10 microns (p-value: <0.001), the latter were better in all the parameters analyzed. However, no difference was noted in the characteristics of the dermal appendages (p-value: >0.05)., Conclusion: While mucosal margins can be obtained at the usual 5-7 microns, the same thickness is not optimal for skin margins intra-operatively. Frozen sections for the skin margins may be set at 10 microns, to save time, minimize artefacts, and for better readability.

Background: The perineum is typically injured at the first vaginal birth. The application of a cold compress to the perineal repair site can reduce pain; however, the effect usually dissipates after a couple of hours. Repeated applications may be needed for sustained analgesia. However, the medium-term effect of repeated applications of cold compress on the perineal repair site on the recovery of sexual function and perineal healing is not known., Objective: This study aimed to evaluate repeated applications of cold vs room temperature (placebo control) compress to the repaired primiparous perineum on pain upon movement., Study Design: A randomized controlled trial was conducted in a university hospital in Malaysia from May 2022 to February 2023. A total of 224 women with a repaired episiotomy or spontaneous second-degree tear sustained at normal delivery were randomized as follows: 113 to frozen gel pack and 111 to room temperature gel pack, as wound compress. The compress was applied to the perineal repair site at 3 timepoints: immediately after repair, and at 4 and 8 hours after delivery, for 20 minutes at each application. The primary outcomes were pain during movement at 12 and 24 hours after delivery, scored using the 0 to 10 numerical rating scale. The secondary outcomes include duration of hospital stay; analgesic consumption; recovery and functional metrics of reestablishing flatus, mobilization, and urination, breastfeeding; maternal satisfaction with the allocated compress; and after hospital discharge for up to 6 weeks after birth through telephone interview, analgesic consumption, perineal pain, resumption of vaginal sex, and women's perception of perineal wound healing., Results: The median (interquartile range) of pain at movement scores were 4 (4-5) vs 5 (4-5) (P=.018) at 12 hours and 2 (1-3) vs 2 (2-3) (P=.173) at 24 hours after birth for cold vs room temperature compress, respectively. Maternal satisfaction scores were 8 (7-9) vs 7 (6-8) (P=.119), oral analgesic for perineal pain while at the postnatal ward was taken by 94 of 113 (83.2%) vs 85 of 109 (78.0%) (relative risk, 1.07; 95% confidence interval, 0.94-1.21), and time to the first satisfactory breastfeeding episode was 11.6 (7.9-15.5) vs 13.0 (8.0-20.7) hours (P=.303) for cold vs room temperature compress, respectively. At 2 weeks telephone follow-up, analgesic intake and perineal pain were not different. At 6 weeks, analgesic intake, perineal pain, resumption of vaginal sex, exclusive breastfeeding, and maternal perception of perineal healing were not different., Conclusion: Intermittent cold compress in the first 8 hours to the repaired perineum reduces pain at 12 hours but the effect attenuates by 24 hours. Maternal satisfaction with their allocated compress was not different. There was no suggestion of harm or benefit on the other secondary outcomes., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Purpose: To determine whether embryo cryopreservation is associated with a difference in maternal serum analyte levels in singleton and twin pregnancies conceived via in vitro fertilization (IVF)., Methods: This was a retrospective cohort study of singleton and twin pregnancies conceived via IVF from a university health system from 01/2014 to 09/2019. Patients with available first and second trimester serum analyte data were included and analyzed separately. Multiple of the median (MoM) values for free β-human chorionic gonadotropin (β-hCG), pregnancy-associated plasma protein A, alpha-fetoprotein (AFP), Inhibin A, and unconjugated estriol (uE3) were compared between two groups: pregnancies conceived after the transfer of fresh embryos versus pregnancies conceived after the transfer of frozen-thawed embryos. Multiple linear regression of log MoM values with F test was performed to adjust for potential confounders., Results: For singletons, fresh embryos were associated with a lower median first trimester free β-hCG (1.00 MoM vs. 1.14 MoM; parameter estimate [PE] 0.90, 95% CI 0.82-0.99, p = .03) compared to frozen-thawed embryos. Fresh embryos were also associated with a lower median second trimester uE3 (0.93 MoM vs. 1.05 MoM; PE 0.88, CI 0.83-0.95, p = .0004) and AFP (1.02 MoM vs. 1.19 MoM; PE 0.91, CI 0.84-0.99, p = .02) compared to frozen-thawed embryos in singletons. There were no significant differences between median first and second trimester serum analytes in twin pregnancies compared between the two groups., Conclusion: Singleton pregnancies derived from fresh embryos had lower first (free β-hCG) and second (uE3 and AFP) trimester analytes compared to frozen-thawed embryos. Twin pregnancies demonstrated no difference between the groups., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Donor sperm treatment is advised to be performed with frozen-thawed donor semen. A disadvantage of frozen-thawed semen is lower pregnancy rates compared to inseminations with fresh semen. Semen parameters affect ongoing pregnancy rates in intracervical inseminations with frozen-thawed donor semen. In an attempt to translate this into clinical relevance, cohort studies have tried to find cut-off values for semen parameters after thawing for intracervical insemination, but these studies assessed only one semen parameter per study, thereby overlooking the intricate interplay between all semen parameters. We performed a retrospective cohort study and tried to calculate thresholds for all semen parameters that lead to the best possible ongoing pregnancy rates in intracervical insemination with frozen-thawed donor semen. Between April 1999 and December 2015, data from 1,186 women who underwent 7,103 cycles of intracervical insemination with donor semen from 129 sperm donors were available for analysis. Our results showed that total motility and total motile count (TMC) after thawing were associated with ongoing pregnancy rate. The best possible ongoing pregnancy chances after intracervical insemination were obtained at a total motility of ≥20% and a total motile count (TMC) of ≥8 × 10 6 after thawing.

Competing Interests: Conflict of interestThe authors have no conflicts of interest to declare.

Background: A precise determination of time since death plays a major role in forensic routine. Currently available techniques for estimating the postmortem interval (PMI) are restricted to specific time periods or cannot be applied for individual case-specific reasons. During recent years, it has been repeatedly demonstrated that Western blot analysis of postmortem muscle protein degradation can substantially contribute to overcome these limitations in cases with different background. Enabling to delimit time points at which certain marker proteins undergo distinct degradation events, the method has become a reasonable new tool for PMI delimitation under various forensic scenarios. However, additional research is yet required to improve our understanding of protein decomposition and how it is affected by intrinsic and extrinsic factors. Since there are temperature limits for proteolysis, and investigators are confronted with frozen corpses, investigation of the effects of freezing and thawing on postmortem protein decomposition in the muscle tissue is an important objective to firmly establish the new method. It is also important because freezing is often the only practical means to intermittently preserve tissue samples from both true cases and animal model research., Methods: Sets of dismembered pig hind limbs, either freshly detached non-frozen, or thawed after 4 months of freeze-storage (n = 6 each), were left to decompose under controlled conditions at 30 °C for 7 days and 10 days, respectively. Samples of the M. biceps femoris were regularly collected at predefined time points. All samples were processed via SDS-PAGE and Western blotting to identify the degradation patterns of previously characterized muscle proteins., Results: Western blots show that the proteins degrade predictably over time in precise patterns that are largely unaffected by the freeze-and-thaw process. Investigated proteins showed complete degradation of the native protein band, partly giving rise to degradation products present in distinct time phases of the decomposition process., Conclusion: This study provides substantial new information from a porcine model to assess the degree of bias that freezing and thawing induces on postmortem degradation of skeletal muscle proteins. Results support that a freeze-thaw cycle with prolonged storage in frozen state has no significant impact on the decomposition behavior. This will help to equip the protein degradation-based method for PMI determination with a robust applicability in the normal forensic setting., (© 2023. The Author(s).)

In microbiome studies, fecal and oral samples are stored and processed in different ways, which could affect the observed microbiome composition. In this study, we compared storage and processing methods applied to samples prior to DNA extraction to determine how each affected microbial community diversity as assessed by 16S rRNA gene sequencing. We collected dental swabs, saliva, and fecal samples from 10 individuals, with three technical replicates per condition. We assessed four methods of storing and processing fecal samples prior to DNA extraction. We also compared different fractions of thawed saliva and dental samples to fresh samples. We found that lyophilized fecal samples, fresh whole saliva samples, and the supernatant fraction of thawed dental samples had the highest levels of alpha diversity. The supernatant fraction of thawed saliva samples had the second highest evenness compared to fresh saliva samples. Then, we investigated the differences in observed community composition at the domain and phylum levels and identified the amplicon sequence variants (ASVs) that significantly differed in relative abundance between the conditions. Lyophilized fecal samples had a greater prevalence of Archaea as well as a greater ratio of Firmicutes to Bacteroidetes compared to the other conditions. Our results provide practical considerations not only for the selection of storage and processing methods but also for comparing results across studies. Differences in processing and storage methods could be a confounding factor influencing the presence, absence, or differential abundance of microbes reported in conflicting studies., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Superdock, Zhang and Poole.)

Glass wool column filtration (GWCF) selects human, bull, boar, dog and buffalo spermatozoa, but reports in the horse are scarce. Single-layer colloid centrifugation with Androcoll-E™ is currently the standard procedure to select good-quality equine sperm. This study was designed to assess GWCF (50 and 75 mg columns; GWCF-50 and GWCF-75, respectively) efficacy to select good-quality sperm from fresh and frozen-thawed equine semen, and to compare its performance with Androcoll-E™ colloid centrifugation. Percentage total motile (TM), progressively motile (PM), morphologically normal (MN), osmotically competent (HOS+) and acrosome-intact/osmotically competent (AI/HOS+) sperm were determined. In studies done with fresh semen samples (n = 17), suspensions subjected to GWCF-50 showed an improvement (p < .05) in PM and HOS+ sperm after selection. With GWCF-75, an increase (p < .05) in PM, MN and HOS+ sperm was observed. Results with GWCF were comparable or better than with Androcoll-E™ selection. Sperm recovery was similar between procedures for all semen parameters. Total sperm count recovery was lower after GWCF-75 (GWCF-50 = 60.0; GWCF-75 = 51.0; Androcoll-E™ = 76.0 million sperm; median; p = .013), but results on total progressive sperm count were similar (GWCF-50 = 23.0; GWCF-75 = 27.0; Androcoll-E™ = 24.0 million sperm; median; p = .3850). Using frozen-thawed semen samples (n = 16), an improvement (p < .05) in TM, PM, NM, HOS+ and AI/HOS+ sperm was observed in GWCF-75 filtrates. Results were comparable to Androcoll-E™ centrifugation, except HOS+ that increased (p < .05) only after GWCF-75. Recovery was comparable for all parameters in frozen samples. GWCF is a simple and low-cost procedure that selects equine sperm with a quality comparable to colloid centrifugation with Androcoll-E™., (© 2023 Wiley-VCH GmbH. Published by John Wiley & Sons Ltd.)

In this study, the effects of freezing yak milk at -20 °C and -40 °C for 30, 90 and 180 days on the fermentation characteristics and storage quality of the corresponding yogurt were discussed. The results showed that, compared with that of yogurt made from fresh yak milk, the lactic acid bacteria (LAB) growth and acid production rate of the yogurt in the -20 °C group decreased at 90 d. The water-holding capacity, viscosity and hardness decreased during storage, and a sour taste was prominent, while no significant changes were observed in the -40 °C group. At 180 d of freezing, the post-acidification of the yogurt in the -20 °C and -40 °C groups increased after 21 d of storage. Compared with the -40 °C group, the -20 °C group showed a significant decrease in LAB counts, a decrease in pH value to 3.63-3.80 and poor texture and sensory quality.

Reactive Oxygen Species (ROS), primarily produced by cellular metabolism, are highly reactive molecules that modify cellular compounds. During sperm preparation in Assisted Reproductive Techniques (ARTs), intrinsic and extrinsic sources of ROS can impact spermatozoa's oxidative status. The modification of the media with compounds that enhance sperm quality characteristics is of great significance. The current study investigated the effect of pterostilbene, a phenolic compound, on bovine sperm quality. Cryopreserved spermatozoa from six bulls were thawed, supplemented with pterostilbene (0, 10 μΜ, 25 μΜ) and incubated for 60 min and 240 min. Spermatozoa were analyzed in terms of motility, viability, acrosomal status and intracellular concentration of superoxide anion in each time point. The incubation of spermatozoa with 25 μΜ pterostilbene resulted in the preservation of quality parameters through superoxide anion mitigation, while its presence in capacitating conditions resulted in higher percentage of acrosome-reacted spermatozoa. The results of the present study indicate that the addition of pterostilbene prevents oxidative insult to spermatozoa and preserves the sperm quality parameters.

Background: The in vitro stability assessment is essential for investigating the diagnostic accuracy of pleural biomarkers. This study aimed to investigate the long-term stability of pleural fluid carcinoembryonic antigen (CEA) at -80°C to -70°C. In addition, we analyzed the effects of frozen storage on the diagnostic accuracy of CEA for malignant pleural effusion (MPE)., Methods: Pleural fluid CEA of participants in two prospective cohorts were stored at -80°C to -70°C for 1-3 years. The CEA level in the stored specimen was measured with an immunoassay, and its level in the fresh specimen was extracted from medical records. The Bland-Altman method, Passing-Bablok regression, and Deming regression were used to analyze the agreement of CEA between the fresh and frozen pleural fluid. In addition, we used receiver operating characteristic (ROC) curves to evaluate the diagnostic accuracy of CEA in the fresh and frozen specimens for MPE., Results: A total of 210 participants were enrolled. The median CEA levels in frozen and fresh pleural fluid specimens were similar (frozen, 2.32 ng/mL; fresh, 2.59 ng/mL; p < 0.01). The slopes and intercepts in the Passing-Bablok regression (intercept 0.01, slope 1.04) and Deming regression (intercept 0.65; slope 1.00) were not statistically significant (p > 0.05 for all). No significant difference was observed between the area under the ROC curves of CEA in the fresh and frozen specimens (p > 0.05 for all)., Conclusion: Pleural fluid CEA is seemingly stable when stored at -80°C to -70°C for 1-3 years. Frozen storage does not significantly affect the diagnostic accuracy of CEA for MPE., (© 2023 The Authors. Thoracic Cancer published by China Lung Oncology Group and John Wiley & Sons Australia, Ltd.)

Aims and Objectives  Gynecological neoplasms are among the most common cancers in female population of India and worldwide. Various new advances have been made to diagnose gynecological pathologies which include imprint cytology and frozen sections in addition to the histopathological techniques. The aim of the study was to assess the diagnostic accuracy of intraoperative diagnostic procedure with gold standard histopathology. Method  The study included 50 patients who had suspected gynecological neoplasm in a time period of 12 months. Their intraoperative diagnosis was made on frozen section and imprint cytology to rule out benign or malignant lesions and compared with gold standard histopathology. Results  Our study concludes that maximum cases of female genital tract neoplasms belonged to the age group of 19 to 76 years, with 60% cases in postmenopausal age group. Overall diagnosis of 62, 52, and 76% malignancies were made on imprint cytology, frozen section, and histopathological examination, respectively. Conclusion  The study concludes that diagnostic accuracy by intraoperative imprint cytology is higher (80%) than frozen section (76%). True positive cases were maximally reported by histopathology. True negative and false positive cases were equally reported by both frozen and imprint cytology. False negative cases were reported by frozen section more than by imprint cytology. The kappa statistical value was lesser in frozen versus histopathology and more in imprint versus histopathology. The sensitivity, specificity, positive predictive value, and negative predictive value for imprint cytology were 77.5, 90.0, 96.9, and 50%, respectively, whereas for frozen section, it was 72.5, 90.0, 96.7, and 45.0%, respectively., Competing Interests: Conflict of Interest None declared., (The Indian Association of Laboratory Physicians. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. ( https://creativecommons.org/licenses/by-nc-nd/4.0/ ).)

Introduction: Frozen sperm utilization might negatively impact cycle outcomes in animals, implicating cryopreservation-induced sperm damage. However, in vitro fertilization and intrauterine insemination (IUI) in human studies are inconclusive., Methods: This study is a retrospective review of 5,335 IUI [± ovarian stimulation (OS)] cycles from a large academic fertility center. Cycles were stratified based on the utilization of frozen ( FROZEN , n  = 1,871) instead of fresh ejaculated sperm ( FRESH , n  = 3,464). Main outcomes included human chorionic gonadotropin (HCG) positivity, clinical pregnancy (CP), and spontaneous abortion (SAB) rates. Secondary outcome was live birth (LB) rate. Odds ratios (OR) for all outcomes were calculated utilizing logistic regression and adjusted (adjOR) for maternal age, day-3 FSH, and OS regimen. Stratified analysis was performed based on OS subtype [ gonadotropins ; oral medications ( OM ): clomiphene citrate and letrozole; and unstimulated/natural ]. Time to pregnancy and cumulative pregnancy rates were also calculated. Further subanalyses were performed limited to either the first cycle only or to the partner's sperm only, after excluding female factor infertility, and after stratification by female age (<30, 30-35, and >35 years old)., Results: Overall, HCG positivity and CP were lower in the FROZEN compared to the FRESH group (12.2% vs. 15.6%, p  < 0.001; 9.4% vs. 13.0%, p  < 0.001, respectively), which persisted only among OM cycles after stratification (9.9% vs. 14.2% HCG positivity, p  = 0.030; 8.1% vs. 11.8% CP, p  = 0.041). Among all cycles, adjOR (95% CI) for HCG positivity and CP were 0.75 (0.56-1.02) and 0.77 (0.57-1.03), respectively, ref: FRESH. In OM cycles, adjOR (95% CI) for HCG positivity [0.55 (0.30-0.99)] and CP [0.49 (0.25-0.95), ref. : FRESH ] favored the FRESH group but showed no differences among gonadotropin and natural cycles. SAB odds did not differ between groups among OM and natural cycles but were lower in the FROZEN group among gonadotropin cycles [adjOR (95% CI): 0.13 (0.02-0.98), ref. : FRESH ]. There were no differences in CP and SAB in the performed subanalyses (limited to first cycles or partner's sperm only, after excluding female factors, or after stratification according to female age). Nevertheless, time to conception was slightly longer in the FROZEN compared to the FRESH group (3.84 vs. 2.58 cycles, p  < 0.001). No significant differences were present in LB and cumulative pregnancy results, other than in the subgroup of natural cycles, where higher LB odds [adjOR (95% CI): 1.08 (1.05-1.12)] and higher cumulative pregnancy rate (34% vs. 15%, p  = 0.002) were noted in the FROZEN compared to the FRESH group., Conclusion: Overall, clinical outcomes did not differ significantly between frozen and fresh sperm IUI cycles, although specific subgroups might benefit from fresh sperm utilization., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Cherouveim, Vagios, Hammer, Fitz, Jiang, Dimitriadis, Sacha, James, Bormann and Souter.)

Sodium hypochlorite is generally used as a disinfectant in washing of freshwater fishes where the safety aspect of health is of concern. Although plant-based essential oils and synthetic chemical agents have been applied, they might contain toxic substances, are expensive and can cause undesirable quality. This research aims to fill the knowledge gap necessary to validate Citrus aurantium juice as a disinfecting agent for preserving striped catfish steaks at -20 °C for 28 days. Fifty (50) ppm sodium hypochlorite was used as a commercial disinfectant (control). The results showed that a negative color characteristic (higher a* and increased b*) was found in the control but not in striped catfish steaks immersed in C. aurantium juice (TM) on days 14 and 28. No significant differences were found in the peroxide value among the treatments on days 14 and 28 ( P > 0.05). A lower accumulation of trichloroacetic acid soluble peptides was detected in TM but not in control, while total volatile basic nitrogen of all treatments was up to standard of fish quality during storage. Contrastingly, the total viable count of both treatments increased to >7.0 log CFU/g on day 28 which did not meet the edible limit of standard for freshwater fishes. The spoilage microbial community was observed on days 0 and 28 of storage which showed a decrease in relative abundance of Acinetobacter, Pseudomonas, Brochothrix, Lactococcus, Carnobacterium, Psychrobacter , and Vagococcus as found in TM on day 28, when compared to the control. Thus, these results implied that C. aurantium juice could replace sodium hypochlorite as an alternative disinfecting agent to control the microbiological spoilage and physico-chemical quality of striped catfish steaks., Competing Interests: The authors declare that they have no competing interests., (© 2023 Dabsantai and Mahidsanan.)

Tooth Shell Technique (TST) with the use of autologous dentin has proven to be a suitable method of grafting in the context of lateral ridge augmentation. This present feasibility study aimed to retrospectively evaluate the preservation by lyophilization of processed dentin. Thus, the frozen stored processed dentin matrix (FST: 19 patients with 26 implants) was re-examined with that of processed teeth used immediately after extraction (IUT: 23 patients with 32 implants). Parameters of biological complications, horizontal hard tissue loss, osseointegration, and buccal lamella integrity were used for evaluation. For complications, the observation period was 5 months. Only one graft was lost (IUT group). In the area of minor complications, without the loss of an implant or augmentation, there were two cases of wound dehiscence and one case with inflammation and suppuration (IUT: n = 3, FST: n = 0). Osseointegration and integrity of the buccal lamella were present in all implants without exception. Statistically, there was no difference between the groups studied for the mean resorption of the crestal width and the buccal lamella. Results of this study show that prepared autologous dentin preserved with a conventional freezer had no disadvantage compared to immediately use autologous dentin in terms of complications and graft resorption in the context of TST., Competing Interests: The authors declare no conflict of interest.

Introduction: Patients with early-stage breast cancer currently undergo sentinel lymph node dissection to evaluate the axillary region. Frozen tissue blocks are evaluated intra operatively and paraffin-embedded samples are studied postoperatively. We explored whether sentinel lymph node dissection adequately reflected axillary involvement (as revealed by the paraffin blocks) in patients with early-stage breast cancer; we sought to avoid axillary dissection., Methods: The agreement/non-agreement rates between the results of axillary ultrasonography and biopsy, sentinel lymph node and axillary dissections, and frozen and paraffin block results, were retrospectively analyzed for 200 patients with early-stage breast cancer. The positive predictive values and accuracies were recorded in those who were positive on both ultrasonography and biopsy. The negative predictive values were calculated for doubly negative cases., Results: The frozen and paraffin block results disagreed in 19 (9.5 %) cases and agreed in 181 (90.5 %). The frozen block and dissection results differed in five of 38 patients who underwent axillary dissection (AD) (one patient did not undergo AD); the results were in agreement in 32. Of the 19 block-disagreement cases, 16 were in the non-neoadjuvant chemotherapy (NAC) group and three in the NAC treatment group. Clinically, the negative predictive values of the frozen and paraffin block data were 80 % in patients lacking axillary involvement., Conclusion: Paraffin block evaluations only (thus, without frozen block examinations) of early-stage breast cancer lymph nodes seem to be sufficient to guide treatment. Also, a thorough clinical examination (with ultrasonography and axillary biopsy) reduces the dissection rate and the associated functional impairments., Competing Interests: Conflict of interest The authors declare no conflict of interests., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Mozzarella cheese was industrially frozen (-18 °C), stored for up to six months, tempered at 4 °C for one or three weeks and the structure and functionality compared to cheese stored at 4 °C and cheese aged at 4 °C for four weeks prior to freezing. When combined with ageing or tempering, the slow industrial freezing minimised changes to the protein network as detected by confocal microscopy and arrested proteolysis. Cheese functionality improved with three weeks of tempering, with properties similar to cheese refrigerated for one month, potentially due to increased proteolysis and protein rehydration. Frozen storage induced β-sheet and β-turn structures, as detected by S-FTIR microspectroscopy, with longer tempering leading to structural recovery in the cheese. This study indicates the proteolysis and functionality of frozen cheese can be optimised with tempering time. It also provides new insights into heat transfer during the industrial freezing and tempering of cheese., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Frozen shoulder (FS) is a common name for shoulder movement limitation with different degrees of shoulder rigidity and pain. It is characterized by varying developmental courses, different levels of shoulder movement limitation, and background ambiguity due to the multiplicity of its causative factors. Systemic inflammatory cytokines monitoring and restraining is easy to apply, fast to conduct, and needs lower costs compared to invasive methods for frozen shoulder stage evaluation and early controlling of its progress to the stage that necessitates surgical intervention. The aim of this review was to assess the recent findings concerning the role of cytokines in FS pathogenesis and the possibility of preventing or controlling their progress through targeting these cytokines by the new drugs candidates, such as hyaluronan (HA), botulinum toxin type A (BoNT A), Tetrandrine, tumor necrosis factor-stimulated gene-6 (TSG-6), and cannabidiol. Searching the PubMed site, we encountered out of 1608 records, from which 16 original studies were included for the quantitative construction of this systematic review screening of the recent studies to investigate the different FS pathogenic pathways. Most of the scenarios are centered around the inflammatory and fibrotic process triggered by synovial and capsular fibroblast stimulation. This mechanism depends mainly on alarmins cytokines, including thymic stromal lymphopoietin (TSLP), interleukin-33 (IL-33), and interleukin-25 (IL-25), with the stimulation of interleukin-1 α (IL-1α), interleukin-1 β (IL-1β), tumor necrosis alpha (TNF-α), cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) in a joint capsule. Different pathways of transforming growth factor- β (TGF-β) stimulation, resulting in overexpression of the fibrotic factors as tenascin C (TNC), fibronectin 1, collagen I (COL 1) and collagen III (COL III), and matrix metalloproteinases (MMPs) in the capsular or synovial/capsular fibroblasts. The overall investigation of these studies led us to conclude that the new drug candidates proved their efficiency in controlling the common pathogenesis of the inflammatory and fibrotic pathways of frozen shoulder and therefore represent a prospect for easy and early controlling and efficiently treating this serious disease., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Alghamdi et al.)

The purpose of this experimentation was to study the evolution of the quality of two types of blast-frozen ready-to-(h)eat meals, tortellini and a vegetable soup, during a short shelf life of 70 days. The analyses, performed in order to identify any variations resulting either from the freezing process or from the subsequent storage, carried out at the temperatures of -30 °C and -18 °C, respectively, examined the consistency of the tortellini and the soup, the acidity and the peroxide value of the oil extracted from them, the phenols and carotenoids present in the soup, the volatile compounds in the tortellini and the soup, and a sensory analysis of both products. The results showed that, during the 70 days of shelf life, there was no variation in the texture of the tortellini, but there were changes in the consistency of the soup, which decreased as the days of storage went on. Furthermore, statistically significant increases ( p < 0.05) in the acidity and in the peroxide value of the oil of the soup were observed during the storage period; however, no statistically significant difference ( p > 0.05) in the peroxide value of the oil of the tortellini was found. Moreover, no quantitative changes were observed in the phenolic compounds and carotenoids in the soup or in the volatile substances of either product. Finally, the sensory analysis confirmed, together with the chemical data, that the blast-freezing process adopted was suitable to maintain the good quality of these fresh meals, even if some technical modifications (in particular, lower freezing temperatures) should be adopted to improve the final quality of the products.

Background  Ovarian neoplasm is the third most common malignancy in Indian women. Intraoperative diagnosis becomes the critical guiding tool for the surgeons to take the decisions on the extent of surgery specially when preserving fertility has to be considered. Aims and Objective  The aim of this study is to evaluate the concordance of intraoperative diagnosis of frozen section (IFS) of ovarian epithelial neoplasm at our institute and to review and discuss the diagnostic pitfalls along with the review of literature. Materials and Methods  Data were archived from departmental record and the detailed clinical data of the patients were retrieved from hospital record system. The discordant cases were reviewed again in an attempt to address the pitfalls. Statistical Analysis  Diagnostic accuracy, sensitivity, specificity, and positive and negative predictive value of IFS of ovarian neoplasm were analyzed. Results  The overall frozen section diagnosis was concordant with final histopathology in 36 out of 44 cases (81%). The sensitivity of IFS diagnosis was found to be 100% for benign and borderline tumors, whereas 88.9% for malignant epithelial tumors, but the correctness of diagnosis is high only for benign and malignant tumors (high positive predictive value) in compared with borderline ones. The diagnostic pitfalls were identified individually in discordant cases. Conclusion  An accurate interpretation of IFS in ovarian epithelial malignancy can be achieved in benign and malignant cases, but limited in borderline tumors. Awareness of the artifacts and the limitations in mind and the IFS diagnosis can be of great help for proper management of the ovarian neoplasm., Competing Interests: Conflict of Interest None declared., (MedIntel Services Pvt Ltd. This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. ( https://creativecommons.org/licenses/by-nc-nd/4.0/ ).)

The effect of freezing raw meat on the quality characteristics of beef jerky was evaluated in the present study. Jerky was made using different types of raw beef (fresh, frozen, and frozen-thawed) with different curing times (6 h and 12 h). Frozen-thawed beef had a lower moisture content than fresh or frozen beef due to higher exudate loss (P < 0.05). Jerky made using frozen and frozen-thawed beef showed lower drying yield and higher shear force than jerky prepared using fresh beef (P < 0.05). Freezing raw beef decreased the fat content and increased the redness, yellowness, chroma, and hue values of jerky (P < 0.05). The microstructure of beef jerky was showed to increase the deformation and contraction of muscle fibers due to freezing. Longer curing times increased the moisture content of jerky made using frozen meat (P < 0.05). Jerky made using frozen or frozen-thawed meat was tough due to excessive fat and moisture loss., Competing Interests: Declaration of Competing Interest The authors declare no potential conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Introduction: Telepathology (TP) allows for remote slide review with performance comparable to traditional light microscopy. Use of TP in the intraoperative setting allows for faster turnaround and greater user convenience by obviating the physical presence of the attending pathologist. We sought to perform a practical validation of an intraoperative TP system using the Leica Aperio LV1 scanner in tandem with Zoom teleconferencing software., Methods: A validation was performed in accordance with recommendations from CAP/ASCP, using a retrospectively identified sample of surgical pathology cases with a 1 year washout period. Only cases with frozen-final concordance were included. Validators underwent training in the operation of the instrument and conferencing interface, then reviewed the blinded slide set annotated with clinical information. Validator diagnoses were compared to original diagnoses for concordance., Results: 60 slides were chosen for inclusion. 8 validators completed the slide review, each requiring 2 h. The validation was completed in 2 weeks. Overall concordance was 96.4%. Intraobserver concordance was 97.3%. No major technical hurdles were encountered., Conclusion: Validation of the intraoperative TP system was completed rapidly and with high concordance, comparable to traditional light microscopy. Institutional teleconferencing implementation driven by the COVID pandemic facilitated ease of adoption., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Background: The Netherlands Armed Forces (NLAF) are using -80°C deep-frozen thrombocyte concentrate (DTC) since 2001. The aim of this study is to investigate the effect of storage duration and alterations in production/measurement techniques on DTC quality. It is expected that DTC quality is unaffected by storage duration and in compliance with the European guidelines for fresh and cryopreserved platelets., Study Design and Methods: Pre-freeze and post-thaw product platelet content and recovery were collected to analyze the effects of dimethyl sulfoxide (DMSO) type, duration of frozen storage (DMSO-1 max 12 years and DMSO-2 frozen DTC max 4 years at -80°C) and type of plasma used to suspend DTC. Coagulation characteristics of thawed DTC, plasma and supernatant of DTC (2× 2500 G) were measured with Kaolin thromboelastography (TEG) and phospholipid (PPL) activity assay., Results: Platelet content and recovery of DTC is ±10%-15% lower in short-stored products and remained stable when stored beyond 0.5 years. Thawed DTC (n = 1724) were compliant to the European guidelines (98.1% post-thaw product recovery ≥50% from original product, 98.3% ≥200 × 10 9 platelets/unit). Compared to DMSO-1, products frozen with DMSO-2 showed ±8% reduced thaw-freeze recovery, a higher TEG clot strength (MA 58 [6] vs. 64 [8] mm) and same ±11 s PPL clotting time. The use of cold-stored thawed plasma instead of fresh thawed plasma did not influence product recovery or TEG-MA., Discussion: Regardless of alterations, product quality was in compliance with European guidelines and unaffected by storage duration up to 12 years of -80°C frozen storage., (© 2022 The Authors. Transfusion published by Wiley Periodicals LLC on behalf of AABB.)

Competing Interests: None disclosed.

Extruded noodles made from whole Tartary buckwheat are widely known as healthy staple foods, while the treatment of fresh noodles after extrusion is crucial. The difference in sensory and functional quality between frozen noodles (FTBN) and hot air-dried noodles (DTBN) was investigated in this study. The results showed a shorter optimum cooking time (FTBN of 7 min vs. DTBN of 17 min), higher hardness (8656.99 g vs. 5502.98 g), and less cooking loss (5.85% vs. 21.88%) of noodles treated by freezing rather than hot air drying, which corresponded to better sensory quality (an overall acceptance of 7.90 points vs. 5.20 points). These effects on FTBN were attributed to its higher ratio of bound water than DTBN based on the Low-Field Nuclear Magnetic Resonance results and more pores of internal structure in noodles based on the Scanning Electron Microscopy results. The uniform water distribution in FTBN promoted a higher recrystallization (relative crystallinity of FTBN 26.47% vs. DTBN 16.48%) and retrogradation (degree of retrogradation of FTBN 34.67% vs. DTBN 26.98%) of starch than DTBN, strengthening the stability of starch gel after noodle extrusion. FTBN also avoided the loss of flavonoids and retained better antioxidant capacity than DTBN. Therefore, frozen treatment is feasible to maintain the same quality as freshly extruded noodles made from whole Tartary buckwheat. It displays significant commercial potential for gluten-free noodle production to maximize the health benefit of the whole grain, as well as economic benefits since it also meets the sensory quality requirements of consumers.

Between 2016 and 2021, the Canadian Food Inspection Agency (CFIA) collected 4218 samples of fresh and frozen berries (blackberries, blueberries, raspberries, strawberries and mixed berries) and pomegranate arils at retail across 11 major cities in Canada and tested these samples for the presence of norovirus GI, norovirus GII and hepatitis A virus RNA. The purpose of this testing was to provide information on the prevalence of these viruses in berries and pomegranate arils on the Canadian marketplace. Of the 926 fresh fruit samples tested, norovirus GI RNA was detected in one raspberry sample and norovirus GII RNA was detected in one strawberry sample. Of the 3292 frozen fruit samples tested, norovirus GI RNA was detected in one blackberry sample, one raspberry sample and one strawberry sample, and norovirus GII RNA was detected in one blueberry sample, three raspberry samples, four strawberry samples, one pomegranate arils sample and one mixed berry sample. None of the fresh or frozen fruit samples tested positive for hepatitis A virus RNA. No statistically significant associations were observed between the prevalence of viral RNA in samples of fresh and frozen fruit, between the prevalence of viral RNA in samples of domestic and imported fruit or between the prevalence of viral RNA in samples of specific fruit types. Overall, the prevalence of norovirus GI and GII RNA together in fresh and frozen fruit samples in Canada was 0.36 %. The results of this study may be used to refine surveillance programs for norovirus and hepatitis A virus in fresh and frozen berries and pomegranate arils, e.g. by adapting the commodities tested and/or the numbers of planned samples to better target these hazards. This information may also be used to inform other Government of Canada approaches to better understand the controls associated norovirus and hepatitis A virus in fresh and frozen berries and pomegranate arils., (Crown Copyright © 2022. Published by Elsevier B.V. All rights reserved.)

Sorbets are healthy ice cream alternatives and desired frozen desserts by vegetarians and vegans. This study focuses on assessing the effects of sorbet recipe alteration through the addition of different percentages of Zizyphus jujuba powder. Stevia rebaudiana was used as a sugar substituent. A control sample and five variables were developed. Peaches from the "Laskava" (native Bulgarian) variety were used as the main ingredient. The new sorbet matrices were characterized based on their moisture and ash content, overrun, melting behavior, melting rate, water holding capacity, pH, nutritional data, soluble solids content, titratable acidity, vitamin C content, total phenolic content and antioxidant activity. The microbial load and CIELAB color of the sorbet alternatives was also evaluated. A sensory evaluation revealed the most preferred variant. Panelists evaluated the appearance ( n = 6), aroma ( n = 5), flavor ( n = 5), mouthfeel ( n = 7), and aftertaste ( n = 5). The soluble solids content varied from 17.50 to 33.03%, the ash content from 0.36 to 5.21%, the moisture content from 63.77 to 80.21%. The studied sorbet matrices have an overrun in the range from 8.11 to 12.32%. Results showed that a potential for the development of peach sorbet matrices with added value and a reasonable consumer acceptability exists. Further research can perfect the recipe and provide a reference for other frozen desserts.

Embryo cryopreservation has been an integral part of ART for close to 40 years and vitrification has boosted overall ART efficacy and safety. Recently, there has been a vivid scientific discussion on whether elective cryopreservation of all embryos (freeze-all) should be pursued for most patients, with a fresh embryo transfer taking place only in selected cases. In terms of efficacy, the available evidence suggests that the freeze-all strategy leads to higher live birth rates after the first embryo transfer compared to the conventional strategy in high responders, while there is no difference in normal responders. There is no evidence to suggest that the freeze-all strategy is inferior to the conventional strategy of fresh transfer when comparing cumulative live birth rates using data from all available randomized controlled trials. The incidence of ovarian hyperstimulation syndrome is significantly reduced in the freeze-all policy. However, regarding obstetric complications and neonatal outcomes, the evidence suggests that each strategy is associated with certain risks and, therefore, there is no approach that could be unequivocally accepted as safer. Similarly, limited evidence does not support the notion that patients would be universally against freeze-all owing to the inevitable delay in pregnancy achievement. Finally, the cost-effectiveness of freeze-all is likely to vary in different settings and there have been studies supporting that this policy can be, under certain conditions, cost-effective. Adoption of the freeze-all policy can also allow for more flexible treatment strategies that have the potential to increase efficacy, reduce cost and make treatment easier for patients and clinics. Importantly, freeze-all does not require the use of any experimental technologies, further training of personnel or the costly acquisition of new equipment. For these reasons, transitioning to the freeze-all policy for most patients appears to be the next logical step in ART., (© The Author(s) 2022. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Aim: During several local COVID-19 outbreaks in China in 2020, SARS-CoV-2 or its RNA was isolated or detected from frozen food or packages, revealing the lack of effective disinfection measures in the frozen food chain and risk of transmission. We explored the possibility that disinfectant plus antifreeze could be delivered as thermal fog to realize effective disinfection at subzero temperatures., Methods and Results: We selected two disinfectant-antifreeze combinations, didecyl dimethyl ammonium bromide (DDAB) - propylene glycol (PPG) and peracetic acid (PAA) - triethylene glycol (TEG), and each combination is used with a custom-optimized thermal fogging machine. The two fogs were tested in -20°C freezer warehouses for their disinfection efficacy against a coronavirus porcine epidemic diarrhoea virus (PEDV) field strain, a swine influenza virus (SIV) field strain, and three indicator bacteria, Escherichia coli, Staphylococcus aureus and Bacillus subtilis endospores. At -20°C, the DDAB-PPG or PAA-TEG thermal fogs settle within 3.5 to 4.5 h and effectively inactivated PEDV with median tissue culture infective dose of 10 -3.5 0.1 ml -1 and SIV-H1N1 with hemagglutination titre of 2 6  ml -1 within 15-60 min. DDAB-PPG could inactivate S. aureus and E. coli vegetative cells (10 6  cfu ml -1 ) within 15-60 min but not effective on B. subtilis spores, while PAA-TEG could disinfect B. subtilis spores more effectively than for S. aureus and E. coli., Conclusions: We showed that a practical subzero temperature disinfection technology was effective in killing enveloped viruses and vegetative bacteria or bacterial spores. DDAB-PPG or PAA-TEG thermal fogging may be a practical technology for cold-chain disinfection., Significance and Impact of the Study: This subzero temperature disinfection technology could help to meet the urgent public health need of environmental disinfection in frozen food logistics against pandemic and other potential pathogens and to enhance national and international biosecurity., (© 2021 The Society for Applied Microbiology.)

Few food waste interventions focus on drivers distinct to particular food groups, such as seafood. Given suggestive evidence that seafood may be wasted at exceptionally high rates, and given its environmental, economic and nutritional value, this research provides insights into seafood-specific consumer food waste interventions. We performed three complementary sub-studies to examine consumer and retailer views regarding seafood waste and frozen seafood as well as perceptions of an intervention providing chef-created recipes to promote cooking frozen seafood without defrosting. The findings indicated an openness to a direct-from-frozen intervention among many consumers and retailers, and suggested seven potential barriers to adoption, along with ways to address them. Underlying the potential for this intervention, and more broadly contributing to addressing consumer seafood waste, the research formed the basis of a new "4 Ps" concept model to characterize the drivers of discarded seafood: proficiency, perceptions/knowledge, perishability, and planning/convenience. These factors shape waste through pathways that include behavioral protocols; taste preferences; waste-prevention efforts; and food safety concerns, precautions, and errors. This research suggested the benefit of testing a larger-scale direct-from-frozen intervention using insights from the concept model and, more broadly, the benefits of exploring approaches to food waste prevention rooted in specific food groups.

Background: Fresh stem cell exosomes are usually obtained and reused in the same individual. It cannot be kept viable for a long period of time regardless of the lengthy preparation time. Freezing is typically used to preserve the viability of perishable materials and increase their lifetime. Regrettably, normal freezing of biomaterials leads to cell damage. Therefore, a cryoprotectant can save the cells from the conventional cryodamage. Sodium carboxymethylcellulose (NA-CMC) is a powdery substance that is used to manufacture bio-safe hydrofilm gels because of its high viscosity, cytocompatibility, and nonallergenic nature. Materials and Methods:  Sterile CMC hydrogel was prepared, part of which was loaded with exosomal solution derived from MSCs. The gel was kept at -20°C for preservation. Two bilateral full-thickness circular skin wounds of 2-cm diameter were created on the back of experimental dogs. The wounds were at least 2.5 cm apart. Treatment started 24 hours after wound creation. Group I received CMC gel solely, whereas group II received frozen CMC exosomal gel. The gel was applied 4 times, a single application per day with 1- day interval. Results: Clinically, the frozen exosomal gel significantly promoted wound healing with no scaring. Histologically, enhanced dermal fibroblasts and organized collagen deposition were seen in the treated group. Conclusion: CMC proved to be an efficient cryoprotectant and a suitable vehicle for exosomes. Deep freezing was proven to conserve the viability, extended the preservation, and facilitated the usage of exosomal gel. This technique of preserved cell-free therapy is inexpensive, time-saving, and proficient and seems suitable for treating cutaneous wounds., (Copyright © 2021 Tehran University of Medical Sciences.)

The present study evaluated an enzyme strategy for eliminating the gliadin in the flour in order to produce part-baked (PB) frozen bread for celiac patients. At first, tissue transglutaminase with lysine methyl ester transamidated the gliadin and hydrolyses gliadin protein. The deamidated dough was used for producing the PB bread and then stored as the frozen storage at - 18 °C for 15 days, followed by investigating physicochemical, rheological, and sensory properties. The SDS-PAGE result demonstrated that transamidating wheat flour with a tissue transglutaminase and L-lysine methyl ester break down the gliadin protein. The PB frozen bread with the absence of gliadin had lower specific volume, porosity, firmness, and color index ( P  < 0.05) but adding 0.8% guar gum could improve these factors and recompense the absence of gliadin ( P  < 0.05). The PB frozen bread with 0.8% guar gum had physicochemical properties such as fresh bread which produced with untreated wheat flour ( P  < 0.05)., (© Association of Food Scientists & Technologists (India) 2020.)

Background: Spontaneous ovulation during a natural menstrual cycle is frequently used for timing frozen embryo transfer (FET). Nevertheless, it remains unclear whether or not women should receive luteal phase support (LPS) following natural cycle frozen embryo transfer (NC-FET)., Objective and Rationale: The aim of this systematic review and meta-analysis was to study whether the administration of LPS improves the reproductive outcome following NC-FET., Search Methods: We conducted a systematic search of the literature published in Medline/PubMed, Embase and the Cochrane Library, from January 2000 until December 2020. We included all original English, peer-reviewed articles, irrespective of the study design. The search strategy included keywords related to NC-FET and luteal phase support. Studies reporting the results of artificial or stimulated FET cycles were excluded., Outcomes: Our systematic search generated 416 records. After screening, eight studies were included in the review and seven studies were included in the meta-analysis. Two studies (n = 858) used hCG and six studies (n = 1507) used progesterone for luteal support. Four studies were randomised controlled trials (RCTs), whereas the other four were historic cohort studies. In a meta-analysis using a random effects model, hCG administration for LPS did not increase the clinical pregnancy rate (CPR) (two studies, odds ratio (OR) 0.85, 95% CI 0.64-1.14). On the other hand, progesterone LPS was associated with a higher CPR (five studies, OR 1.48, 95% CI 1.14-1.94), and a higher live birth rate (LBR) (three studies, OR 1.67, 95% CI 1.19-2.36). The association between progesterone LPS and the LBR remained significant after excluding non-randomised studies., Wider Implications: The available evidence indicates that progesterone administration for LPS is beneficial following NC-FET. There is no evidence to support the administration of hCG for LPS in these cases. Additional large RCTs are necessary to improve the quality of evidence and validate our findings., (© The Author(s) 2021. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

Purpose: Response to anti-PD-L1/PD-1 immunotherapy correlates with PD-L1 expression in breast cancer. However, the prevalence of PD-L1 positive breast cancer is variable, which could be due to differences in the population/cohort of patients tested or the preservation/detection technology used. To investigate this variability, we examined the effect of two tissue preservation methods on PD-L1 immunohistochemical detection in breast cancer., Methods: We compared PD-L1 expression in patient-matched frozen (FR) and formalin-fixed paraffin-embedded (FFPE) tissues of breast cancer patients. PD-L1 expression was assessed using tumor proportion score (TPS, simply PD-L1 score), and case positivity was determined with PD-L1 score ≥5., Results: In FFPE tissues, PD-L1 was positive in 7-10% of tested patients, depending on the antibody used. In patient-matched FR tissues, the same antibodies showed positive PD-L1 expression in 20-30% of cases. The impact of the antibody tested on the rate of PD-L1 positivity (% of PDL1 positive cases) was minor, as evident in the near perfect concordance between PD-L1 score obtained using the different antibodies whether tested in FR or FFPE tissues. However, there was a systematic drop by an average of 13-20% in the PD-L1 score obtained in FFPE tissues compared to their patient-matched FR tissues., Conclusions: In the tested patient-matched cohort, there was consistently a higher PD-L1 score in FR than FFPE tissues, regardless of the antibody used, demonstrating a significant effect on PD-L1 detection due to the preservation method. These findings should inspire further work to improve the sensitivity of PD-L1 detection and possibly search for more sensitive antibodies in FFPE tissues.

Comet assay, applied to in vitro, in vivo and ex vivo systems, is a quick, simple, and sensitive method for the detection of genotoxicity. In general, fresh whole blood or peripheral blood mononuclear cells (PBMCs) are used in the assay for the determination of DNA damage and repair. In this study, the effects of storage conditions on genotoxicity assessed by Comet assay in human whole blood and lymphocyte samples, were evaluated. Whole blood and lymphocyte samples were stored at 4 °C for 1, 2, 3, 4, 5 and 7 days; at -20 °C for 1 month and at -80 °C for 3, 6 and 12 months. 1% DMSO was used as cryoprotectant. No significant differences in DNA damage were demonstrated in all of the storage conditions and durations, and the results were similar according to the median values (p < 0.05). According to Spearman or Pearson correlations, an important correlation was found between the DNA damage of the fresh samples and the samples which were kept at -80 °C for 6 months with temperature and time (p < 0.01 for Pearson and p < 0.05 for Spearman). The results of this study indicated that blood and lymphocyte samples stored in +4 °C, -20 °C and -80 °C up to 12 months can be used instead of fresh samples especially in human biomonitoring studies., (Copyright © 2021 Elsevier Ltd. All rights reserved.)