Characterization of ovarian tissue oocytes from transgender men reveals poor calcium release and embryo development, which might be overcome by spindle transfer.

STUDY QUESTION: Can spindle transfer (ST) overcome inferior embryonic development of in vitro matured ovarian tissue oocytes (OTO-IVM) originating from testosterone-treated transgender men? SUMMARY ANSWER: ST shows some potential to overcome the embryo developmental arrest observed in OTO-IVM oocytes from transgender men. WHAT IS KNOWN ALREADY: OTO-IVM is being applied as a complementary approach to increase the number of oocytes/embryos available for fertility preservation during ovarian tissue cryopreservation in cancer patients. OTO-IVM has also been proposed for transgender men, although the potential of their oocytes remains poorly investigated. Currently, only one study has examined the ability of OTO-IVM oocytes originating from transgender men to support embryo development, and that study has shown that they exhibit poor potential. STUDY DESIGN, SIZE, DURATION: Both ovaries from 18 transgender men undergoing oophorectomy were collected for the purposes of this study, from November 2020 to September 2022. The patients did not wish to cryopreserve their tissue for fertility preservation and donated their ovaries for research. All patients were having testosterone treatment at the time of oophorectomy and some of them were also having menses inhibition treatment. PARTICIPANTS/MATERIALS, SETTING, METHODS: Sibling ovaries were collected in either cold or warm medium, to identify the most optimal collection temperature. Cumulus oocyte complexes (COCs) from each condition were isolated from the ovarian tissue and matured in vitro for 48 h. The quality of OTO-IVM oocytes was assessed by calcium pattern releasing ability, embryo developmental competence following ICSI, and staining for mitochondrial membrane potential. In vitro matured metaphase I (MI) oocytes, germinal vesicle (GV) oocytes, and in vivo matured oocytes with aggregates of smooth endoplasmic reticulum (SERa) were donated from ovarian stimulated women undergoing infertility treatment and these served as Control oocytes for the study groups. ST was applied to overcome poor oocyte quality. Specifically, enucleated mature Control oocytes served as cytoplasmic recipients of the OTO-IVM spindles from the transgender men. Embryos derived from the different groups were scored and analysed by shallow whole genome sequencing for copy number variations (CNVs). MAIN RESULTS AND THE ROLE OF CHANCE: In total, 331 COCs were collected in the cold condition (OTO-Cold) and 282 were collected in the warm condition (OTO-Warm) from transgender men. The maturation rate was close to 54% for OTO-Cold and 57% for OTO-Warm oocytes. Control oocytes showed a calcium releasing ability of 2.30 AU (n = 39), significantly higher than OTO-Cold (1.47 AU, P = 0.046) oocytes (n = 33) and OTO-Warm (1.03 AU, P = 0.036) oocytes (n = 31); both values of calcium release were similar between the two collection temperatures. Mitochondrial membrane potential did not reveal major differences between Control, OTO-Warm, and OTO-Cold oocytes (P = 0.417). Following ICSI, 59/70 (84.2%) of Control oocytes were fertilized, which was significantly higher compared to 19/47 (40.4%) of OTO-Cold (P < 0.01) and 24/48 (50%) of OTO-Warm oocytes (P < 0.01). In total, 15/59 (25.4%) blastocysts were formed on Day 5 in the Control group, significantly higher than 0/19 (0%) from the OTO-Cold (P = 0.014) and 1/24 (4.1%) in OTO-Warm oocytes (P = 0.026). Application of ST rescued the poor embryo development, by increasing the Day 5 blastocyst rate from 0% (0/19) to 20.6% (6/29) (P = 0.034), similar to that in the ICSI-Control group (25.4%, 15/59). A normal genetic profile was observed in 72.7% (8/11) of OTO-Cold, 72.7% (8/11) of OTO-Warm and 64.7% (11/17) of Control Day 3-Day 5 embryos. After ST was applied for OTO-IVM oocytes, 41.1% (7/17) of the embryos displayed normal genetic patterns, compared to 57.1% (4/7) among ST-Control Day 3-Day 5 embryos. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: Due to the limited access to human oocytes and ovarian tissue, our results should be interpreted with some caution, as only a limited number of human oocytes and embryos could be investigated. WIDER IMPLICATIONS OF THE FINDINGS: The results of this study, clearly indicate that OTO-IVM oocytes originating from transgender patients are of inferior quality, which questions their use for fertility preservation. The poor quality is likely to be related to cytoplasmic factors, supported by the increased blastocyst numbers following application of ST. Future research on OTO-IVM from transgender men should focus on the cytoplasmic content of oocytes or supplementation of media with factors that promote cytoplasmic maturation. A more detailed study on the effect of the length of testosterone treatment is also currently missing for more concrete guidelines and guidance on the fertility options of transgender men. Furthermore, our study suggests a potentially beneficial role of experimental ST in overcoming poor embryo development related to cytoplasmic quality. STUDY FUNDING/COMPETING INTEREST(S): A.C. is a holder of FWO grants (1S80220N and 1S80222N). A.B. is a holder of an FWO grant (1298722N). B.H. and A.V.S. have been awarded with a special BOF (Bijzonder Onderzoeksfonds), GOA (Geconcerteerde onderzoeksacties) and 2018000504 (GOA030-18 BOF) funding. B.H. has additional grants from FWO-Vlaanderen (Flemish Fund for Scientific Research, G051516N and G1507816N) and Ghent University Special Research Fund (Bijzonder Onderzoeksfonds, BOF funding (BOF/STA/202109/005)), and has been receiving unrestricted educational funding from Ferring Pharmaceuticals (Aalst, Belgium). The authors declare that they have no conflict of interest. TRIAL REGISTRATION NUMBER: N/A.

Influência da frequência estimulatória envolvida nos efeitos analgésicos induzidos por eletroacupuntura em cervicalgia tensional / Influence of the stimulating frequency involved in analgesic effects induced by electroacupuncture for neck pain due to muscular tension

Objetivo: Avaliar a influência da frequência estimulatória envolvida na analgesia induzida por eletroacupuntura em cervicalgia. Métodos: Comparou-se o desempenho da analgesia produzida em 2Hz, 100Hz, 1000Hz, 2500Hz e um grupo só com acupuntura, sem estímulo elétrico, avaliado por meio de algometria de pressão, Escala Visual Analógica (EVA) e frequência cardíaca. Utilizou-se um estimulador elétrico microprocessado, com forma de pulso em padrão pulsado, monofásico, retangular, balanceado assimétrico, comfase secundária em exponencial decrescente, com período de estimulação de 4 segundos e repouso de 3 segundos. A amostra contou com 66 voluntários com cervicalgia tensional, idade média de 33,67±9,97 anos, 89,5% do gênero feminino e 10,5% do masculino. Resultados: Não houve diferenças entre os grupos para as variáveis nota atribuída à dor pela EVA e frequência cardíaca, sendo que em todos os grupos houve melhoras analgésicas. No entanto, quando comparado o comportamento antes-depois, por meio da algometria de pressão, para um mesmo indivíduo, dentro de seu próprio grupo, houve vantagens analgésicas para o uso de 2500Hz (p=0,006 para a base da região occiptal; p=0,003 para o trapézio direito; e p=0,013 para o trapézio esquerdo), seguido de 100Hz(p=0,035, p=0,016 e p=0,038, para as mesmas regiões, respectivamente). Conclusão: Recomenda-se preferencialmente a aplicação de 2500Hz e 100Hz em eletroacupuntura para analgesia em cervicalgia tensional.

Objective: To assess the influence of the stimulating frequency involved in analgesia induced by electroacupuncture for neck pain. Methods: The performance of the analgesia produced by 2Hz, 100Hz, 1000Hz and 2500Hz was compared with a group withacupuncture alone (without electrical stimulation), by means of pressure algometry, a visual analog scale (VAS) and heart rate. We used an electrical stimulator with a microprocessor yielding standard, single-phase, rectangular and asymmetrical balanced pulsed waveforms with a secondary phase decreasing exponentially. Stimulation periods were 4s, and resting periods were 3s. The sample included 66 volunteers with neck pain due to muscular tension, mean age 33.67±9.97 years, 89.5% female and 10.5% male. Results: There were no differences between the groups regarding the variables of degree of pain (according to the VAS) and heart rate, and all groups presented analgesic improvement. However, when comparing pressure algometry findings for the same individual before and after the treatment, within the same group, we found analgesic advantages in using 2500Hz (p=0.006 for the base of the occipital region, p=0.003 for the right trapezius and p=0.013 for the left trapezius), followed by 100Hz (p=0.035, p=0.016 and p=0.038 for thesame regions, respectively). Conclusion: We preferentially recommend 2500Hz and 100Hz applications of electroacupuncture for analgesia of neck pain due to muscular tension.

[The use of a multimix as a dietary supplement: study in rats]. / Uso de uma multimistura como suplementação alimentar: Estudo em ratos.

A Multimistura (MM)--sweet cassava (Manihot esculenta Crantz) leaf flour, wheat bran (Tritium aestivum L.), egg shell powder, pumpkin (Cucurbita Spp) and sunflower (Heliantus annus) seed flours--was added to a mixture of Beans, 7% (Phaseolus vulgaris) and Rice, 3% (Oryza sativa) and its effects, were assessed in weanling, male albino (Wistar) rats (n = 60). Animals were divided into 6 groups: groups 1, 2 and 3 were fed beans + rice + multimixture (B + R + MM), beans + rice (B + R) and 10% Casein, respectively; the remaining groups were maintained on a protein-free diet (PFD) for 14 d and then submitted to the same feeding protocol. Microbiological assays were performed in all MM samples. The Coefficient of Digestablity (CD), the Food Efficiency Ratio (FER), Protein Efficiency Ratio (PER), Net Protein Utilization (NPR), serum hemoglobin (Hb) and hematocrit (Ht), carcass total lipids were determined. Rats had their liver, brain, gonads, testes, spleen and left kidney removed for wet dry weights. Liver samples were histologically examined. The Mann-Whitney test was used. The protein content of B + R diet increased slightly after MM addition (0.23 g/100 g). Three out of four MM samples had moulds and yeasts. CD values were 90% and 70% for casein and B + R + MM-fed rats, respectively. The highest values for FER, PER and NPR were seen in the casein-fed rats without protein depletion. The casein-fed group had heavier organs (wet and dry weights) and higher values for carcass fat and serum Hb and Ht. Steatosis was present in both groups, with or without protein depletion. Short or long-term MM consumption, at least under our experimental conditions, had no significant effects on investigated parameters.

Dietary modification of chyle composition in chylothorax.

Nutrition support has played a major role in the treatment of chylothorax, both to prevent malnutrition and to minimize chyle production and flow. This report evaluates chyle composition in a patient with chylothorax who was placed on a low-fat diet, medium-chain triglyceride diet, and total parenteral nutrition in sequence. Both triglyceride content and volume of chyle declined, but drainage persisted, ultimately requiring thoracic duct ligation. The chyle triglyceride while on total parenteral nutrition, which presumably originates from both the intestine and plasma, contained more long-chain unsaturated fatty acids than the circulating serum triglyceride. Of particular interest was the detection of an appreciable amount of medium-chain fatty acids in the chyle triglyceride, constituting 20% of the triglyceride fatty acids when an enteral formulation with medium-chain triglyceride as a sole fat source was administered. The finding of almost threefold more decanoic acid (C10:0) than octanoic acid (C8:0), despite the presence of considerably more octanoic acid in the original diet, suggests that trioctanoin may be a preferable medium-chain triglyceride substrate for the nonsurgical treatment of chylothorax.

Attenuation of the febrile response in guinea pigs by fish oil enriched diets.

The influence of dietary lipid manipulation on the fever response to an injection of murine recombinant Interleukin-1 (rIL-1) in guinea pigs was examined. The animals were fed diets identical except for the lipid source for periods of 5 and 6 wk. In vitro thromboxane B2/B3 (TxB2/B3) production in collagen-stimulated whole blood was also measured. One diet was enriched with menhaden oil, high in the omega-3 series of fatty acids. The other diet contained safflower oil, consisting primarily of the omega-6 fatty acid linoleic acid (74%). Animals fed the fish oil-enriched diet for 6 wk not 5 week had a blunted fever response after rIL-1 injection. This was associated with a 27% increase (p less than 0.05) in total omega-3 fatty acids in plasma phospholipids between weeks 5 and 6 in fish oil-fed animals. Safflower oil-fed animals produced a "normal" febrile response regardless of the duration of feeding. Safflower-fed guinea pigs demonstrated a significant increase in TxB2/B3 production in whole blood after 6 wks that was associated with a 25% increase (p less than 0.05) in plasma phospholipid arachidonic acid levels between weeks 5 and 6. Despite significant reductions in phospholipid linoleic acid in animals fed fish oil, arachidonic acid levels remained unchanged. In the guinea pig model used, long-term menhaden oil feeding can significantly blunt the febrile response induced by exogenous Interleukin-1. Also, a high intake of linoleic acid as seen in safflower oil feeding can significantly increase thromboxane production in stimulated whole blood.(ABSTRACT TRUNCATED AT 250 WORDS)