Effect of IL-1ß on the Development of Spermatogenesis In Vitro in Normal and Busulfan-Treated Immature Mice.

Gonadotoxic agents could impair spermatogenesis and may lead to male infertility. The present study aimed to evaluate the effect of IL-1ß on the development of spermatogenesis from cells isolated from seminiferous tubules (STs) of normal and busulfan-treated immature mice in vitro. Cells were cultured in a 3D in vitro culture system for 5 weeks. We examined the development of cells from the different stages of spermatogenesis by immunofluorescence staining or qPCR analyses. Factors of Sertoli and Leydig cells were examined by qPCR analysis. We showed that busulfan (BU) treatment significantly reduced the expression of testicular IL-1ß in the treated mice compared to the control group (CT). Cultures of cells from normal and busulfan-treated immature mice induced the development of pre-meiotic (Vasa), meiotic (Boule), and post-meiotic (acrosin) cells. However, the percentage of developed Boule and acrosin cells was significantly lower in cultures of busulfan-treated mice compared to normal mice. Adding IL-1ß to both cultures significantly increased the percentages of Vasa, Boule, and acrosin cells compared to their controls. However, the percentage of Boule and acrosin cells was significantly lower from cultures of busulfan-treated mice that were treated with IL-1ß compared to cultures treated with IL-1ß from normal mice. Furthermore, addition of IL-1ß to cultures from normal mice significantly increased only the expression of androgen receptor and transferrin but no other factors of Sertoli cells compared to their CT. However, the addition of IL-1ß to cultures from busulfan-treated mice significantly increased only the expression of androgen-binding protein and the FSH receptor compared to their CT. Adding IL-1ß to cultures of normal mice did not affect the expression of 3ßHSD compared to the CT, but it significantly reduced its expression in cultures from busulfan-treated mice compared to the CT. Our findings demonstrate the development of different stages of spermatogenesis in vitro from busulfan-treated mice and that IL-1ß could potentiate this development in vitro.

Identification of spermatogenesis in individual seminiferous tubules and testicular tissue of adult normal and busulfan-treated mice employing Raman spectroscopy and principal component analysis.

The present study aims to identify spermatogenesis in testicular seminiferous tubules (ST) and testicular tissue of adult normal and busulfan-treated mice utilizing PCA and Raman spectroscopy. Raman measurements were conducted on single tubules and testes samples from adult and immature mice, comparing them with those from busulfan-treated adult mice, with validation through histological examination. The analysis revealed a higher signal variability (30 %-40 % at the peaks), prompting scrutiny of individual Raman spectra as a means of spermatogenesis measurement. However, principal component analysis (PCA) demonstrated significant cluster separation between the ST of mature and immature mice. Similar investigations were performed to compare ST from normal mature mice and those from busulfan-treated (BS-treated) mature mice, revealing substantial separation along PC1 and PC2 for all comparison sets. Additionally, comparing testicular samples from mature and immature mice revealed distinct separation in PCA. The study concludes that the combined approach of PCA and Raman spectroscopy proves to be a noninvasive and potentially valuable method for identifying spermatogenesis in seminiferous tubules and testicular samples.

Radiographic Examination before Dental Extraction from Dentists' Perspective.

Background: It is generally agreed that radiographic examination is important before dental extraction. It provides information about the roots and the surrounding tissues. In terms of practice, it does not seem to be a universally implemented protocol regarding the use of dental radiology before dental extraction. Besides, the type of radiographic technique is not specified. Some references prefer periapical dental radiographs. Others prefer orthopantomography), or even cone beam computed tomography Delpachitra et al. (2021) [1]. In terms of the dental practice, it is not clear whether there is a universally adopted protocol regarding the use of dental radiographs before dental extraction. Aim of the study. To assess dental professionals' perspective toward radiographic examination before conventional dental extraction. Materials and Methods: A Google form questionnaire was circulated to different dental professionals using mainly ResearchGate, in addition to different social media platforms. Results: One hundred and forty-five dentists participated in the questionnaire. The respondents were divided according to the country of current practice: national (Iraqi), regional (Middle Eastern), and international participants. Out of 144 respondents, 51.4% percent of the participants were international, while 40.3% were Iraqis, and 8.3% were from the Middle East. The need for dental radiography in all dental extraction procedures was reported in the majority of responses (n = 86). Only 11 dentists think there is no necessity for radiographic examination before conventional extraction. The chi-square test showed a highly significant relationship between the country of current practice and the need for X-ray examination for conventional dental extraction (P < 0.01). Seventy-six dentists prefer periapical radiographs. Thirty-five preferred orthopantomography. A highly significant relationship was found between the country of practice and the X-ray technique (P < 0.01). Conclusion: The study showed that there is no universally adopted protocol regarding the use of dental radiography before dental extraction. The country of practice appears to govern the dentists' decisions regarding the need for an X-ray and the type of radiography prior to dental extraction. Periapical radiographs for posterior teeth seem to be the preferable choice before dental extraction.

Report of a novel mutation in CRB1 in a Lebanese family presenting retinal dystrophy.

PURPOSE: To identify the genetic basis of a recessive inheritance form of retinal dystrophy (RD) in a Lebanese family. MATERIALS AND METHODS: Clinical data were recorded for five patients of the 14 family members. Genetic linkage was carried out using Affymetrix 250 K Nspl SNP array followed by sequencing. RESULTS: The patients showed variable phenotypes ranging from Leber Congenital Amaurosis (LCA) to progressive forms of Retinitis Pigmentosa (RP). A 46.1 Mb chromosomal region at chromosome 1q23.3-32.2 was identified by homozygosity mapping. This region contained the Crumbs homologue-1, CRB1, a gene responsible for recessive retinal dystrophies. CRB1 is required for photoreceptor morphogenesis, and it has been associated with RP and LCA. Sequencing of CRB1 revealed two mutations: a novel deletion in exon 6 (c.1772_1775delGCAT; p.C591Sfs*28) and a missense mutation in exon 7 (c.2234C > T; p.T745M). CONCLUSION: We report a novel CRB1 mutation in inherited RD in a Lebanese family, and confirm the considerable phenotype heterogeneity that may exist between individuals sharing the same mutations.