A review and statistical analysis to identify and describe relationships between CQAs and CPPs of natural killer cell expansion processes.

Natural killer (NK) cells make only a small fraction of immune cells in the human body, however, play a pivotal role in the fight against cancer by the immune system. They are capable of eliminating abnormal cells via several direct or indirect cytotoxicity pathways in a self-regulating manner, which makes them a favorable choice as a cellular therapy against cancer. Additionally, allogeneic NK cells, unlike other lymphocytes, do not or only minimally cause graft-versus-host diseases opening the door for an off-the-shelf therapy. However, to date, the production of NK cells faces several difficulties, especially because the critical process parameters (CPPs) influencing the critical quality attributes (CQAs) are difficult to identify or correlate. There are numerous different cultivation platforms available, all with own characteristics, benefits and disadvantages that add further difficulty to define CPPs and relate them to CQAs. Our goal in this contribution was to summarize the current knowledge about NK cell expansion CPPs and CQAs, therefore we analyzed the available literature of both dynamic and static culture format experiments in a systematic manner. We present a list of the identified CQAs and CPPs and discuss the role of each CPP in the regulation of the CQAs. Furthermore, we could identify potential relationships between certain CPPs and CQAs. The findings based on this systematic literature research can be the foundation for meaningful experiments leading to better process understanding and eventually control.

Biomechanical Integrity Score of the Female Pelvic Floor for Stress Urinary Incontinence.

INTRODUCTION AND HYPOTHESIS: This study is aimed at developing and validating a new integral parameter, the Biomechanical Integrity score (BI-score) of the female pelvic floor for stress urinary incontinence conditions. METHODS: A total of 130 subjects were included in the observational cohort study; 70 subjects had normal pelvic floor conditions, and 60 subjects had stress urinary incontinence (SUI). A Vaginal Tactile Imager (VTI) was used to acquire and automatically calculate 52 biomechanical parameters for eight VTI test procedures (probe insertion, elevation, rotation, Valsalva maneuver, voluntary muscle contractions in two planes, relaxation, and reflex contraction). Statistical methods were applied (t test, correlation) to identify the VTI parameters sensitive to the pelvic SUI conditions. RESULTS: Twenty-seven parameters were identified as statistically sensitive to SUI development. They were subdivided into five groups to characterize tissue elasticity (group 1), pelvic support (group 2), pelvic muscle contraction (group 3), involuntary muscle relaxation (group 4), and pelvic muscle mobility (group 5). Every parameter was transformed to its standard deviation units using the dataset for normal pelvic conditions, similar to the T-score for bone density. Linear combinations with specified weights led to the composition of five component parameters for groups 1-5 and to the BI-score in standard deviation units. The p value for the BI-score has p = 4.0 × 10-28 for SUI versus normal conditions. CONCLUSIONS: Quantitative transformations of the pelvic tissues, support structures, and functions under diseased conditions may be studied with the SUI BI-score in future research and clinical applications.

A randomized controlled pilot trial to assess the effectiveness of a specially formulated food supplement and pelvic floor muscle training in women with stress-predominant urinary incontinence.

BACKGROUND: Pelvic floor muscle training (PFMT) is the first-line treatment approach for stress urinary incontinence. Creatine and leucine have been shown to improve muscle function. Our aim was to assess the effectiveness of a food supplement and PFMT in women with stress-predominant urinary incontinence. METHODS: Women with stress-predominant urinary incontinence were randomized in 1:1 ratio to receive daily oral supplementation for six weeks with either a food supplement (treatment group) or placebo (control group). Both groups were instructed to perform standardized daily PFMT. The primary outcome was the Urogenital Distress Inventory Short Form (UDI-6) score. Secondary outcomes were the Incontinence Impact Questionnaire (IIQ-7) score, Patient's Global Impression of Severity (PGI-S), and Biomechanical Integrity score (BI-score) measured by Vaginal Tactile Imager. To have a power of 80% and a significance level of 5% to detect a decrease of 16 points in the UDI-6 score, a sample size of 32 was needed, with 16 patients in each arm of our trial. RESULTS: Sixteen women in the control group and sixteen in the treatment group completed the trial. Between-group analysis revealed no significant differences between the control and treatment group except for mean change (delta) in vaginal squeeze pressure [(cmH2O, mean ± SD), 5 ± 12 vs. 15 ± 15, P = 0.04] and mean change (delta) in PGI-S score [(mean ± SD), -0.2 ± 0.9 vs. -0.8 ± 0.8, P = 0.04]. Within-group analysis showed that UDI-6 and IIQ-7 scores improved significantly from baseline to six weeks in the treatment group but not in the control group [UDI-6 score (mean ± SD) 45 ± 21 vs. 29 ± 21, P = 0.02; 43 ± 18 vs. 33 ± 26, P = 0.22] [IIQ-7 score (mean ± SD) 50 ± 30 vs. 30 ± 21, P = 0.01; 48 ± 23 vs.40 ± 28, P = 0.36]. PGI-S scores only improved in the treatment group from baseline to six weeks after treatment [PGI-S score (mean ± SD) 3.1 ± 0.8 vs. 2.3 ± 0.8, P = 0.0001]. BI-score, on average, improved significantly in the treatment and control group as well [SD unit, mean, from - 1.06 to -0.58, P = 0.001; from - 0.66 to -0.42, P = 0.04]. CONCLUSIONS: Women with stress-predominant urinary incontinence receiving a specially formulated supplement in addition to daily PFMT for six weeks had significantly improved urinary symptoms (decrease in UDI-6 score and IIQ-7) and BI-score compared to their baseline. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT05358769. 27/04/2022.

Correlation between the female pelvic floor biomechanical parameters and the severity of stress urinary incontinence.

BACKGROUND: Stress urinary incontinence (SUI) is a common condition that requires proper evaluation to select a personalized therapy. Vaginal Tactile Imaging (VTI) is a novel method to assess the biomechanical parameters of the pelvic floor. METHODS: Women with SUI were enrolled in this cross-sectional study. Participants completed the Medical, Epidemiologic, and Social Aspects of Aging (MESA) questionnaire and the Patient Global Impression of Severity Question (PGI-S) and underwent a VTI examination. Based on the MESA and PGI-S questionnaires, participants were divided into mild, moderate, and severe SUI groups. Fifty-two biomechanical parameters of the pelvic floor were measured by VTI and compared between the groups (mild vs. moderate and severe). SUI Score and Index were calculated from the MESA questionnaire. Pearson correlation was used to determine the strength of association between selected VTI parameters and the MESA SUI Index and MESA SUI Score. RESULTS: Thirty-one women were enrolled into the study. Significant differences were observed in the VTI parameters 16, 22-24, 38, 39 when the difference between mild and severe subgroups of SUI based on the PGI-S score was examined. Parameter 16 refers to the maximum gradient at the perineal body, parameter 22-24 refers to the pressure response of the tissues behind the vaginal walls, and parameter 38, 39 refers the maximum pressure change and value on the right side at voluntary muscle contraction. VTI parameter 49, describing the displacement of the maximum pressure peak in the anterior compartment, showed a significant difference between the mild SUI and the moderate-severe SUI according to the MESA SUI score (mean ± SD 14.06 ± 5.16 vs. 7.54 ± 7.46, P = 0.04). The MESA SUI Index and SUI Score displayed a positive correlation concerning VTI parameters 4 (the maximum value of the posterior gradient) and 27 (the displacement of the maximum pressure peak in the anterior compartment) (VTI4 vs. MESA SUI Index r = 0.373, P = 0.039; VTI4 vs. MESA SUI Score r = 0.376, P = 0.037; VTI27 vs. MESA SUI Index r = 0.366, P = 0.043; VTI27 vs. MESA SUI Score r = 0.363, P = 0.044). CONCLUSIONS: Female pelvic floor biomechanical parameters, as measured by VTI, correlate significantly with the severity of SUI and may help guide therapeutic decisions.

Evaluation of zinc and copper levels in vaginal tissues and whole blood: correlation with age.

BACKGROUND: Zinc and copper are essential trace elements and play a crucial role in the homeostasis of connective tissues. In this study, we aimed to define zinc and copper levels in the vaginal tissue and establish whether a correlation exists between the zinc and copper levels either or both in whole blood or vaginal tissue samples and whether the finding correlates with the age of the patient or at least with her menopausal status. METHODS: We collected whole blood and vaginal tissue samples from 32 women and measured their zinc and copper levels by inductively coupled plasma optical emission spectrometry. We have performed Student's t test to evaluate the differences in the mean levels of trace elements and multiple regression to evaluate the association between vaginal tissue zinc/copper levels and age, menopausal status, number of vaginal deliveries, and zinc/copper blood levels. RESULTS: Zinc levels were significantly higher in both the vaginal tissues and whole blood samples than copper levels (p < 0.01). In the vaginal tissue samples, a strong positive correlation could be detected between zinc and copper levels (r = 0.82, p < 0.01). In the vaginal tissue, a negative correlation was found for zinc and copper levels with the age of women (r = - 0.27, p = 0.04 and r = - 0.56, p < 0.01). Multiple linear regression model (age, menopausal status, vaginal delivery and copper/zinc blood levels) showed that only age remained a significant predictor for zinc and copper vaginal tissues levels (p = 0.03, 95% CI - 2.28 to - 0.06; p = 0.004, 95% CI - 1.76 to - 0.34). CONCLUSIONS: Zinc and copper levels in the vaginal tissue decline with age. Out of the examined variables (age, menopausal status, vaginal delivery, and copper/zinc levels), only age is a significant predictor of vaginal zinc/copper levels.

Fractional CO2 laser treatment effect on cervicovaginal lavage zinc and copper levels: a prospective cohort study.

BACKGROUND: The basic principle of vaginal laser therapy is the rejuvenation of the affected tissue. Zinc and copper are essential nutritional trace elements and have a key role in connective tissue homeostasis. We aimed to investigate the effect of vaginal, fractional CO2 laser treatment on cervicovaginal lavage (CVL) zinc and copper levels. METHODS: Twenty-nine postmenopausal women with symptoms of vaginal dryness were enrolled in our prospective cohort study. Three treatments with MonaLisa Touch CO2 laser system were performed four weeks apart. At each treatment CVL was collected, Vaginal Health Index (VHI) was obtained, and Visual Analog Scale (VAS) for vaginal dryness was assigned by patients. Zinc and copper concentrations were measured with optical emission spectrometry before each treatment and six weeks after the 3rd treatment. RESULTS: The VHI scores significantly improved after each laser treatment (mean ± SD VHI score, 13.03 ± 4.49 before vs. 15.55 ± 4.35 after the 1st, 17.79 ± 4.57 after the 2nd and 19.38 ± 4.39 after the 3rd treatment, P < 0.01). Similarly, VAS scores reflected improvement (mean ± SD VAS score 6.59 ± 2.86 before vs. 4.17 ± 2.86 after the 1st, 2.45 ± 2.43 after the 2nd and 1.41 ± 1.94 after the 3rd treatment, P < 0.01). CVL zinc levels were significantly higher compared to copper levels (0.06 ± 0.04 vs. 0.006 ± 0.006 mg/L, P < 0.01) at baseline. While copper levels remained the same through treatments, the CVL zinc level was significantly higher after the second laser treatment compared to the baseline. CONCLUSIONS: Fractional CO2 laser treatment of the vagina impacts CVL zinc and copper levels differently. While CVL copper levels were not different after each laser treatment, zinc levels were significantly higher after the second treatment before returning to baseline values.

Microbial technologies for biotherapeutics production: Key tools for advanced biopharmaceutical process development and control.

Current trends in the biopharmaceutical market such as the diversification of therapies as well as the increasing time-to-market pressure will trigger the rethinking of bioprocess development and production approaches. Thereby, the importance of development time and manufacturing costs will increase, especially for microbial production. In the present review, we investigate three technological approaches which, to our opinion, will play a key role in the future of biopharmaceutical production. The first cornerstone of process development is the generation and effective utilization of platform knowledge. Building processes on well understood microbial and technological platforms allows to accelerate early-stage bioprocess development and to better condense this knowledge into multi-purpose technologies and applicable mathematical models. Second, the application of verified scale down systems and in silico models for process design and characterization will reduce the required number of large scale batches before dossier submission. Third, the broader availability of mathematical process models and the improvement of process analytical technologies will increase the applicability and acceptance of advanced control and process automation in the manufacturing scale. This will reduce process failure rates and subsequently cost of goods. Along these three aspects we give an overview of recently developed key tools and their potential integration into bioprocess development strategies.

The Effect of Vaginal Microablative Fractional CO2 Laser Treatment on Vaginal Cytology.

BACKGROUND AND OBJECTIVES: Most recently vaginal laser treatment was introduced as a new option for women with genitourinary syndrome of menopause, vaginal dryness. Our objective was to assess the effects of intravaginal CO2 laser treatment on vaginal cytology. STUDY DESIGN/MATERIALS AND METHODS: Fifty-two women with symptoms of vaginal dryness were enrolled and underwent vaginal laser treatment using a fractional CO2 laser. Patients received three vaginal laser treatments 4 weeks apart. Vaginal cytology was obtained before the first treatment and 4 weeks after each additional treatment. Vaginal dryness was assessed by using a Visual Analog Scale (VAS). RESULTS: Out of the 52 women enrolled, 34 were in menopause. Postmenopausal women had significantly lower vaginal maturation values (VMV) compared with premenopausal women at the baseline visit (mean ± standard deviation [SD], 42 ± 23 vs. 68 ± 13, P < 0.01). The vaginal dryness VAS was higher (worse) in postmenopausal women compared with premenopausal cases (mean ± SD, 5.7 ± 4 vs. 2.4 ± 3, P < 0.01). The VMV did not change significantly over time after vaginal laser treatment. However vaginal dryness VAS improved significantly after each treatment. Both in the premenopausal and postmenopausal groups, vaginal dryness scores improved significantly from baseline after the three treatments (postmenopausal 5.7 ± 4 vs. 1.6 ± 2.5, P < 0.01 and premenopausal 2.4 ± 3 vs. 0.2 ± 0.5, P < 0.01). Those patients who had improvement in VMV had significantly better (lower) dryness VAS compared with those women without an improvement in VMV after the three treatments (mean ± SD, 0.3 ± 0.8 vs. 1.6 ± 2.6, P = 0.04). CONCLUSIONS: Vaginal dryness VAS improved significantly in a cohort of premenopausal and postmenopausal women undergoing vaginal CO2 laser treatment despite no significant change in vaginal cytology. Lasers Surg. Med. © 2020 Wiley Periodicals, Inc.

The Effect of Fractional CO2 Laser Treatment on the Symptoms of Pelvic Floor Dysfunctions: Pelvic Floor Distress Inventory-20 Questionnaire.

BACKGROUND AND OBJECTIVES: To assess the improvement on pelvic floor distress (PFD)-related urogenital symptoms using validated questionnaires after intravaginal CO2 laser treatment. STUDY DESIGN/MATERIALS AND METHODS: Forty postmenopausal women with genitourinary symptoms of menopause (GSM) were enrolled into this prospective cohort study and underwent vaginal laser treatment using MonaLisa Touch® fractional CO 2 laser system. Patients received three vaginal laser treatments with 360° probe 4 weeks apart. A three-component Pelvic Floor Distress Inventory (PFDI-20) validated questionnaire was filled out by each patient before each session and 4 weeks after the final treatment. Wilcoxon rank sum test was used to compare the before and after treatment scores. RESULTS: Pelvic Organ Prolapse Distress Inventory (POPDI-6) scores were not significantly different after the first treatment compared with baseline (mean ± standard deviation [SD], 21 ± 18 vs. 17 ± 15, P = 0.44). However, each subsequent treatment resulted in further, statistically significant improvement in symptom scores (14 ± 15, P = 0.03 and 13 ± 13, P = 0.01, after the second and third treatments, respectively). Similarly, Urinary Distress Inventory (UDI-6) scores were not significantly different after the first laser treatment (mean ± SD, 36 ± 25 vs. 29 ± 23, P = 0.36). After the second and third treatments there were significant improvement in the standardized scores (24 ± 20, P = 0.03 and 22 ± 21, P = 0.01). Colorectal-Anal Distress Inventory (CRADI-8) scores did not change significantly after three laser treatments. CONCLUSIONS: Three sessions of microablative fractional CO2 vaginal laser treatment significantly improves patient reported urinary and pelvic organ prolapse symptoms. Lasers Surg. Med. © 2019 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

On-line glucose monitoring by near infrared spectroscopy during the scale up steps of mammalian cell cultivation process development.

NIR spectroscopy is a non-destructive tool for in-situ, on-line bioprocess monitoring. One of its most frequent applications is the determination of metabolites during cultivation, especially glucose. Previous studies have usually investigated the applicability of Near Infrared (NIR) spectroscopy at one bioreactor scale but the effect of scale up was not explored. In this study, the complete scale up from shake flask (1 L) through 20 L, 100 L and 1000 L up to 5000 L bioreactor volume level was monitored with on-line NIR spectroscopy. The differences between runs and scales were examined using principal component analysis. The bioreactor runs were relatively similar regardless of scales but the shake flasks differed strongly from bioreactor runs. The glucose concentration throughout five 5000 L scale bioreactor runs were predicted by partial least squares regression models that were based on pre-processed spectra of bioreactor runs and combinations of them. The model that produced the lowest error of prediction (4.18 mM on a 29 mM concentration range) for all five runs in the prediction set was based on the combination of 20 L and 100 L data. This result demonstrated the capabilities and the limitations of an NIR system for glucose monitoring in mammalian cell cultivations.

Association between pelvic organ prolapse types and levator-urethra gap as measured by 3D transperineal ultrasound.

OBJECTIVES: To evaluate the association between pelvic organ prolapse (POP) types and levator-urethra gap (LUG) as measured by 3-dimensional transperineal tomographic ultrasound. METHODS: A retrospective study was carried out on 98 women with symptomatic POP. Three-dimensional transperineal tomographic ultrasound images and POP quantification coordinates were reviewed. Each vaginal compartment was staged for the degree of prolapse, and total number of involved compartments identified. LUG was measured on 3-dimensional tomographic sonograms as the distance between the center of the urethra and the levator insertion bilaterally. Based on prior studies, an abnormal LUG of 25 mm or greater indicated levator avulsion. The LUG and the presence or absence of unilateral/bilateral avulsions was analyzed with reference to the clinical diagnosis of prolapse (single versus multicompartment, and mild [stage II] versus severe [stage III-IV]). Generalized logit models were used to evaluate the association between avulsion and prolapse type and stage. RESULTS: The LUG was substantially larger in women with multicompartment compared to single-compartment POP (28.9 ± 4.1 mm versus 22.7 ± 4.1 mm, P < .01). Similarly, LUG was substantially larger in women with severe (stage III-IV) compared to mild (stage II) POP (28.8 ± 4.7 mm versus 23.3 ± 4.5 mm, P < .01). Women with severe prolapse were 32 times more likely than women with mild prolapse to have bilateral levator avulsion. Those with POP involving all 3 vaginal compartments were 76 times more likely than single-compartment POP to have bilateral levator avulsions. CONCLUSIONS: Bilateral levator ani avulsion as diagnosed by LUG measurements of 25 mm or greater at rest is associated with multicompartment, severe prolapse.

The Effects of Heat Exposure on Vaginal Smooth Muscle Cells: Elastin and Collagen Production.

AIMS: To evaluate smooth muscle cells (SMCs) proliferation and elastin and collagen production after heat exposure (65°C). METHODS: Samples were taken from the anterior vaginal wall, SMCs were cultured, and heated to 65°C for 30 and 60 s. Cell proliferation was assessed; tropoelastin and collagen production was measured. RESULTS: Heat does not affect SMC proliferation at 65°C neither at 30 nor at 60 s. Surface-deposited elastin level was significantly increased after heat exposure (mean ± SD, 30 s 155 ± 5% of control [p < 0.01] and 60 s 516 ± 40% of control [p < 0.01]). Tropoelastin levels in the culture media were significantly lower after 60 s of heat exposure (mean ± SD, 30 s 102 ± 5% of control [p = ns] and 60 s 70 ± 2% of control [p = 0.04]). Significant increase in surface-deposited collagen production was found (mean ± SD, 30 s 170 ± 6% of control [p < 0.01] and 60 s 123 ± 6% of control [p < 0.01]), but no such elevation was measured in the media after heat exposure (mean ± SD, 30 s 120 ± 20% of control [p = ns] and 60 s 100 ± 20% of control [p = ns]). CONCLUSION: When SMCs are exposed to heat there is significant elevation in collagen and elastin production. Changes in the composition of the extracellular matrix after heat exposure may contribute to vaginal wall remodeling.

[Efficacy of the percutaneous tibial nerve stimulation in the treatment of lower urinary tract symptoms]. / A percutan nervus tibialis stimuláció szerepe egyes kismedencei kórképek kezelésében.

Percutaneous tibial nerve stimulation (PTNS) is a minimally invasive, safe and well-tolerated neuromodulation technique for the lower urinary tract dysfunctions. PTNS delivers neuromodulation to the pelvic floor through the S2-4 junction of the sacral nerve plexus via the route of the posterior tibial nerve. Using the fine needle electrode insertion above the ankle, the tibial nerve is accessed, which connected to the stimulator. To date despite of its excessive clinical use, PTNS mechanism of action still remains unclear. The technique seems to be an efficacious and safe treatment for overactive bladder syndrome (OAB). It could be recommended according to the Urinary Incontinence Guideline of the European Association of Urology in women who did not have adequate improvement or could not tolerate anti-muscarinic therapy. The success rate is comparable to sacral nerve stimulation in OAB patients. PTNS has been used for fecal incontinence since 2003, however, many of the published studies are of poor quality. PTNS has also been shown to have positive effects on chronic pelvic pain, when the usual therapeutic steps did not result in satisfactory improvement. No major complications are reported in the literature, following PTNS treatment. Orv Hetil. 2018; 159(43): 1735-1740.

The enhanced inhibitory effect of different antitumor agents in self-microemulsifying drug delivery systems on human cervical cancer HeLa cells.

The aim of this study was to develop topical self-microemulsifying drug delivery systems (SMEDDS) containing antitumor agents (bleomycin, cisplatin and ifosfamide) and to investigate their inhibitory potential in SMEDDS on human cervical cancer HeLa cells. The physicochemical properties of cytostatic drug loaded SMEDDS were characterized. The cytotoxicity of main components of SMEDDS was also investigated. Their IC50 values were determined. HeLa cells were treated by different concentrations of cisplatin, bleomycin and ifosfamide alone and in various SMEDDS. The inhibitory effect on cell growth was analyzed by MTT cell viability assay. Inflammation is a driving force that accelerates cancer development. The inhibitory effect of these antitumor agents has also been tested on HeLa cells in the presence of inflammatory mediators (IL-1-ß, TNF-α) as an in vitro model of inflamed human cervix. Significant differences in the cytotoxicity of cytostatic drugs alone and in SMEDDS have been found in a concentration-dependent manner. The self-micro emulsifying system may potentiate the effectiveness of bleomycin, cisplatin and ifosfamide topically. The effect of SMEDDS containing antitumor agents was decreased significantly in the presence of inflammatory mediators. According to our experiments, the optimal SMEDDS formulation is 1:1:2:6:2 ratios of Isopropyl myristate, Capryol 90, Kolliphor RH 40, Cremophor RH40, Transcutol HP and Labrasol. It can be concluded that SMEDDS may increase the inhibitory effect of bleomycin, ifosfamide and cisplatin on human cervical cancer HeLa cells. Inflammation on HeLa cells hinders the effectiveness of SMEDDS containing antitumor agents. Our results might ensure useful data for development of optimal antitumor formulations.

Zinc prevents vaginal candidiasis by inhibiting expression of an inflammatory fungal protein.

Candida causes an estimated half-billion cases of vulvovaginal candidiasis (VVC) every year. VVC is most commonly caused by Candida albicans, which, in this setting, triggers nonprotective neutrophil infiltration, aggressive local inflammation, and symptomatic disease. Despite its prevalence, little is known about the molecular mechanisms underpinning the immunopathology of this fungal infection. In this study, we describe the molecular determinant of VVC immunopathology and a potentially straightforward way to prevent disease. In response to zinc limitation, C. albicans releases a trace mineral binding molecule called Pra1 (pH-regulated antigen). Here, we show that the PRA1 gene is strongly up-regulated during vaginal infections and that its expression positively correlated with proinflammatory cytokine concentrations in women. Genetic deletion of PRA1 prevented vaginal inflammation in mice, and application of a zinc solution down-regulated expression of the gene and also blocked immunopathology. We also show that treatment of women suffering from recurrent VVC with a zinc gel prevented reinfections. We have therefore identified a key mediator of symptomatic VVC, giving us an opportunity to develop a range of preventative measures for combatting this disease.

Lymphocyte expansion in bioreactors: upgrading adoptive cell therapy.

Bioreactors are essential tools for the development of efficient and high-quality cell therapy products. However, their application is far from full potential, holding several challenges when reconciling the complex biology of the cells to be expanded with the need for a manufacturing process that is able to control cell growth and functionality towards therapy affordability and opportunity. In this review, we discuss and compare current bioreactor technologies by performing a systematic analysis of the published data on automated lymphocyte expansion for adoptive cell therapy. We propose a set of requirements for bioreactor design and identify trends on the applicability of these technologies, highlighting the specific challenges and major advancements for each one of the current approaches of expansion along with the opportunities that lie in process intensification. We conclude on the necessity to develop targeted solutions specially tailored for the specific stimulation, supplementation and micro-environmental needs of lymphocytes' cultures, and the benefit of applying knowledge-based tools for process control and predictability.

Application of ultrasound elastography in obstetrics and gynecology / Ultrahang-elasztográfia alkalmazásának lehetoségei a szülészet-nogyógyászatban.

Összefoglaló. Az ultrahang-elasztográfia az elmúlt évek során egyre növekvo figyelmet kapott a lágyszövetek elaszticitásának vizsgálatában. A módszer használatát az teszi szükségessé, hogy egyes, a mechanikai tulajdonságaikban különbözo szövetek hasonló echogenitásúak lehetnek, valamint hogy egy adott szövet megváltozott struktúrája vagy mechanikai tulajdonsága nem minden esetben jár együtt a szövet hagyományos ultrahangképének megváltozásával. Az elmúlt évtizedben a deformációs és a nyírási ultrahang-elasztográfia vált széles körben elérhetové. Ezen új képalkotási technika egyre nagyobb szerepet tölt be a szülészeti-nogyógyászati ultrahang-diagnosztikában is. A nogyógyászatban szerephez juthat az endometriosis és az adenomyosis kimutatásában, valamint a benignus és a malignus cervicalis és ovarialis képletek elkülönítésében. A nogyógyászathoz hasonlóan a szülészetben is jelentos változást hozhat az ultrahang-elasztográfia: alkalmas lehet a szülésindukció sikerességének, a koraszülés bekövetkezésének és a praeeclampsia kialakulásának elorejelzésére. Orv Hetil. 2021; 162(18): 690-695. Summary. Ultrasound elastography has received significant attention for the assessment and measurement of soft tissue elastic properties in recent years. The advantage of ultrasound elastography lies in the fact that two different tissues can share similar echogenicities but may have other mechanical properties or, on the contrary, mechanical abnormalities of a designated tissue do not necessarily go hand in hand with an altered appearance on a conventional ultrasound image. In the last decade, strain and shear-wave elasticity imaging methods have become the most widely available among commercial ultrasound equipments. The importance of this new method expands rapidly also in the field of obstetrics and gynecology. Ultrasound elastography has a promising role in the diagnosis of endometriosis and adenomyosis and helps to differentiate benign and malignant cervical and ovarian lesions. The use in the prediction of the outcome of labor induction and preterm birth, and in the evaluation of preeclampsia are emerging. Orv Hetil. 2021; 162(18): 690-695.

Zinc Transporter 9 (SLC30A9) Expression Is Decreased in the Vaginal Tissues of Menopausal Women.

Our aim was to compare zinc transporter (ZnT/SLC30A, and ZIP/SLC39A) expression between pre- and postmenopausal women in human vaginal tissues. Zinc transporter families are responsible for the maintenance of intracellular zinc concentrations. Zinc has significant effects on the extracellular matrix composition. Vaginal wall biopsies were obtained from seven premenopausal and seven postmenopausal women. mRNA expression of twenty-four zinc transporters was determined by quantitative real-time PCR. Zinc transporter expression at the protein level was assessed by immunohistochemistry. Student's t test and Mann-Whitney U test were used to compare data. ZnT4 and ZnT9 mRNA expression were significantly lower in postmenopausal women compared with premenopausal women (mean ± SD mRNA expression in relative units, 96.43 ± 140.61 vs. 410.59 ± 304.34, p = 0.03 and 0.62 ± 0.39 vs. 1.13 ± 0.31, p = 0.02). In addition, ZIP2, ZIP3, and ZIP6 mRNA expressions were significantly lower in postmenopausal women compared with premenopausal women (mean ± SD mRNA expression in relative units, 1.11 ± 0.61 vs. 2.29 ± 1.20, p = 0.04; 2.32 ± 1.90 vs. 15.82 ± 12.97, p = 0.02 and 1.10 ± 0.80 vs. 5.73 ± 4.72, p = 0.03). ZnT9 protein expression in the stratum spinosum was significantly lower in postmenopausal women (p = 0.012). Zinc transporters were expressed differentially in the vaginal tissues. ZnT9 expression was significantly lower in postmenopausal women compared with premenopausal women.

The effect of oral zinc supplementation on cervicovaginal lavage fluid zinc level.

OBJECTIVES: To investigate the effect of oral zinc supplementation on cervicovaginal lavage fluid (CVL) zinc level in pre and postmenopausal women. STUDY DESIGN: A prospective interventional cohort study was carried out by the enrollment of twelve premenopausal and ten postmenopausal women without significant gynecological conditions. Women received daily oral supplementation with 30 mg of zinc for two weeks. Clinical and demographic variables were stored in a dedicated database. Vaginal Health Index was calculated, and vaginal cytology was obtained. CVL and serum samples were collected in a standardized fashion before and after completion of the oral supplementation. Zinc and copper levels were measured by inductively coupled plasma optical emission spectrometry. Paired t-test was used to compare the before and after treatment results. RESULTS: Serum zinc levels increased significantly both in the pre and postmenopausal women (0.88 ± 0.17 vs. 1.06 ± 0.23, p < 0.01 and 0.83 ± 0.24 vs. 0.96 ± 0.33, p < 0.01) after two weeks of daily oral zinc supplementation. CVL zinc level was significantly higher in the premenopausal group compared to the postmenopausal group before and after supplementation (0.13 ± 0.05 vs. 0.06 ± 0.04, p < 0.01 and 0.10 ± 0.03 vs. 0.05 ± 0.01, p < 0.01). Zinc supplementation had no significant impact on the CVL zinc level in either group. Neither serum nor CVL copper levels were affected by the zinc supplementation. There was no significant correlation between serum and CVL zinc or copper levels. CONCLUSION: Daily oral supplementation with 30 mg of zinc had no significant impact on CVL zinc level despite a significant rise in serum zinc level.

Randomized controlled trial for improved recovery of the pelvic floor after vaginal delivery with a specially formulated postpartum supplement.

OBJECTIVE: To improve pelvic floor recovery after vaginal delivery with daily supplementation of a specially formulated postpartum recovery supplement. METHODS: Within 48 hours of vaginal delivery, primipara women were randomized in a 1:1 ratio to receive daily oral supplementation for 6 weeks with either a combination of regular prenatal vitamin (PNV), leucine (4 g/day), zinc (30 mg/day) and omega-3 fatty acid (900 mg/day) (treatment group), or only a PNV daily (control group). Co-primary outcomes were vaginal squeeze pressure as measured by perineometer and levator muscle injury as measured by transperineal 3-dimensional tomographic ultrasound at 6 weeks postpartum. RESULTS: Twenty-six women in the control group and 27 in the treatment group completed the trial. Weak pelvic floor muscle strength was significantly less frequent in the treatment group compared to the control group at 6 weeks after delivery (28% vs. 58%, P=0.03). Both right and left-sided levator-urethra gap was significantly larger in the control group compared to the treatment group indicating more levator injury being present in the control group at 6 weeks after delivery. Anterior vaginal wall prolapse at or beyond the hymenal ring was significantly more common in the control group compared to the treatment group (19% vs. 0%, P=0.02). Significantly more women reported bothersome bulge symptoms in the control group compared to the treatment group at 6 weeks postpartum (19% vs. 0%, P=0.02). CONCLUSION: Postpartum women who received a specially formulated postpartum recovery supplement had improved recovery of the pelvic floor after vaginal delivery.