Micropterus salmoides). Our results demonstrated that inhibition of mTORC1 signaling promoted autophagy and apoptosis of granulocytes while inhibiting cell proliferation. Moreover, inhibition of the mTORC1 signaling pathway enhanced the phagocytosis capacity of granulocytes. Collectively, our findings revealed the evolutionarily conserved role of the mTORC1 signaling pathway in regulating granulocyte responses, thus providing novel insights into the function of granulocytes in teleost fish.Granulocytes are crucial innate immune cells that have been extensively studied in teleost fish. Studies in mammals have revealed that mechanistic target of rapamycin complex 1 (mTORC1) signaling acts as a significant immune regulatory hub, influencing granulocyte immune function. To investigate whether mTORC1 signaling also regulates the immune function of granulocytes in teleost fish, we established a model of RAPA inhibition of the mTORC1 signaling pathway using granulocytes from largemouth bass ( Granulocytes are essential phagocytes found in various vertebrates, including fish, amphibians, reptiles, birds, and mammals. These cells play a fundamental role in the innate immune defense against various pathogenic microorganisms ,2. WheneThe mechanistic target of rapamycin (mTOR) is a highly conserved protein ~280 kDa) found in all eukaryotes . mTOR fo0 kDa fouMicropterus salmoides) as an early vertebrate model. Our results revealed that RAPA treatment effectively inhibited mTORC1 signaling, as evidenced by the downregulation of S6 and 4EBP1 phosphorylation in granulocytes. Furthermore, the RAPA treatment had a significant impact on several key signaling pathways in granulocytes, including mTOR, cell cycle, apoptosis, autophagy, endocytosis, and phagosome signaling pathways, as well as corresponding changes in cellular physiological phenomena. Notably, the mTORC1 signaling pathway also plays a critical role in granulocyte immune responses, particularly affecting phagocytosis. Our findings provide essential evidence elucidating the mechanism of mTORC1 regulation in granulocyte immune and physiological responses within teleost models. Therefore, this study contributes significantly to our understanding of the mTORC1 signaling pathway and its influence on granulocyte function from an evolutionary perspective, offering crucial insights into the functional evolution of granulocytes in early vertebrates.To investigate whether and how mTORC1 signaling regulates granulocyte immune responses, we utilized largemouth bass  for 30 min inhibited the mTORC1 signaling pathway in granulocytes, which is known to regulate granulocyte function. However, high-dose RAPA exposure did not have any appreciable effects on mTORC2. It has been reported that certain cell lines with RAPA-sensitive AKT/PKB phosphorylation  experience impaired mTORC2 integrity at 1 h after RAPA (100 nM) treatment and almost complete loss of the complex at 24 h . AlthougNext, we proceeded to sequence the transcriptome of granulocytes treated with RAPA for 30 min in vitro. Excitingly, the mTORC1 signaling pathway, the target pathway of RAPA, was successfully enriched, confirming the reliability of the transcriptome results. Moreover, the pathways enriched after RAPA treatment were mainly related to essential cell functions such as cell cycle, apoptosis, autophagy, ribosome biogenesis in eukaryotes, ribosome, DNA replication, and RNA transport. This finding is noteworthy, as similar signaling pathways related to cell activities are generally regulated by the mTORC1 signaling pathway in mammals . Therefobad, bax, caspase 8, and caspase 9 are upregulated during apoptosis [The coordination between cell growth and proliferation is vital to ensure normal cell cycle progression and survival, and conflicting signals may induce cell death or senescence ,49. mTORpoptosis . Moreovepoptosis , and ATGpoptosis . TherefoPhagocytosis by phagocytes is a crucial defense mechanism of the body, enabling the devouring and destruction of particulate matter . In mammMicropterus salmoides) were obtained from a fish farm in E Zhou  and then maintained in a recirculating water system at Huazhong Agricultural University for over 4 weeks before the experiment. The fish (about 120 fish) were kept at 26 °C and fed daily with a commercial diet. Feeding was terminated 48 h prior to the sampling experiment.Healthy largemouth bass  at 10 mg/mL and diluted with PBS before use. The final applied concentration of RAPA in the RAPA group was 100 nM, and as a control, the same volume of DMSO was diluted with PBS in the DMSO group. Subsequently, the 48-well plates were placed in a cell incubator at 26 °C with 5% CO2 for 30 min, and the granulocytes were collected after centrifugation at 1000 g for the next analysis.The largemouth bass (~50 g) were used for in vitro RAPA  treatment studies. Head kidney granulocytes were suspended in DMEM (supplemented with 5% FBS) and divided into 48-well plates with 200 μL per well and a final density of 2 × 10® following the manufacturer’s instructions. PCR products were enriched, quantified, and finally sequenced on the illumina novaseq6000. The clean reads were mapped to the largemouth bass  genome. By using Hisat2’s soft tools, only reads with a perfect match or one mismatch were further analyzed and annotated based on the largemouth bass genome. Differential expression analysis of two groups was performed using DESeq2. The FDR ≤ 0.05 and Fold Change ≥ 2 were set as the thresholds for significantly differential expression. For further analysis of the DEGs, we carried out a KEGG enrichment using KOBAS software [The granulocyte samples of the DMSO group and the RAPA group were sent to Biomarker Technologies Co., Ltd. . Briefly, the RNA was extracted using a TRIzol reagent , and the concentration and integrity of the RNA were checked by NanoDrop 2000  and the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system , respectively. Thereafter, the RNA was used for stranded RNA sequencing library preparation using a VAHTS Universal V6 RNA-seq Library Prep Kit for Illuminasoftware  to ident240/244 (5364), anti-phospho-4EBP1 Thr37/46 (2855) monoclonal rabbit antibody  or anti-β-actin (AC026) monoclonal rabbit antibody  for 1 h on a shaker. After washing four times with PBST (5 min/time), goat anti-rabbit IgG was added and incubated for 45 min at room temperature. After washing four times with PBST (5 min/time), the reaction bands were visualized with the ECL substrate (Bio-Rad Laboratories) and then scanned by the Amersham Imager 600 Imaging System . The densitometry of the target band was analyzed with ImageQuant TL software .To detect the inhibition of the mTORC1 signaling pathway in neutrophils, Western blots were performed. The intracellular proteins were resolved on a 12% SDS-PAGE gel  under reducing conditions. Then the gels were transferred onto a PVDF membrane and blocked in PBS with 8% skim milk . After that, the membrane was incubated with anti-S6 (2217), anti-4EBP1 (9644), anti-phospho-S6 SerTM EDU-647 kit  according to the manufacturer’s instructions. Briefly, to detect the percentage of proliferation granulocytes after RAPA treatment, the head kidney granulocytes were collected after 30 min of RAPA treatment, and 20 μg of 5-ethynyl-2′-deoxyuridine (EdU)  was added to the granulocyte culture media in vitro. After 1 d, the cells were fixed, permeabilized, and stained for EDU. Then, cells were analyzed using a CytoFLEX LX flow cytometer .The proliferation assay was performed with BeyoclickThe apoptosis assay was performed with the AnnexinV-FITC kit  according to the manufacturer’s instructions. Briefly, to detect the effect of RAPA treatment on granulocyte apoptosis, the head kidney granulocytes were collected after 30 min of 100 nM DMSO or RAPA treatment. After washing two times, Annexin-V-FITC was added to the granulocytes, incubated for 15 min at room temperature away from light. After washing two times, PI  was added to the granulocytes, incubated for 15 min at 4 °C, and analyzed using a CytoFLEX LX flow cytometer .2 at 26 °C for 15 min. Cytochalasin D  was added immediately to a final concentration of 5 μM, and the granulocytes were collected by centrifugation, discarding the supernatant. Then, 0.4% Taipan blue was added to quench the fluorescence of the free beads, incubated for 1 min, and then centrifuged to collect the granulocytes. The granulocytes were resuspended and immediately analyzed using a CytoFLEX LX flow cytometer .To detect the effect of RAPA treatment on granulocyte phagocytosis. The phagocytosis assay was performed as described previously with sight modifications . Brieflyt-test and one-way ANOVA. Data were expressed as the mean ± SEM, and p < 0.05 was considered statistically significant.Statistical analyses were conducted by GraphPad Prism 8 software. Data comparisons between two groups were detected by a paired Student’s In conclusion, our study provides compelling evidence for a conserved role of mTORC1 signaling in the function of granulocytes in teleosts. Notably, inhibiting mTORC1 signaling in largemouth bass granulocytes significantly affected cell cycle progression, proliferation, apoptosis, and autophagy. Moreover, our results demonstrate for the first time that mTORC1 signaling plays a crucial regulatory role in the phagocytosis of teleost granulocytes. Given the functional analogy between granulocytes in fish and mammals, our findings suggest that the elucidated mTORC1 signaling pathway plays an evolutionarily conserved role in regulating granulocyte responses, thus providing valuable insights into the function of granulocytes in teleost fish.


Baijiu cellar mud using UA as the sole carbon and energy source. The bacterial strain was identified as Bacillus paramycoides-YC02 through 16S rDNA sequence analysis. Under optimal culture conditions at an initial pH of 7.0 and 38 °C, YC02 completely biodegraded an initial UA concentration of 500 mg/L within 48 h. Furthermore, cell-free extracts of YC02 were found to catalyze and remove UA. These results demonstrate the strong biodegradation ability of YC02 toward UA. To gain further insight into the mechanisms underlying UA biodegradation by YC02, the draft genome of YC02 was sequenced using Illumina HiSeq. Subsequent analysis revealed the presence of gene1779 and gene2008, which encode for riboflavin kinase, flavin mononucleotide adenylyl transferase, and flavin adenine dinucleotide (FAD)-dependent urate hydroxylase. This annotation was based on GO or the KEEG database. These enzymes play a crucial role in the metabolism pathway, converting vitamin B2 to FAD and subsequently converting UA to 5-hydroxyisourate (HIU) with the assistance of FAD. Notably, HIU undergoes a slow non-enzymatic breakdown into 2-oxo-4-hydroxy-4-carboxy-5-ureidoimidazoline (OHCU) and (S)-allantoin. The findings of this study provide valuable insights into the metabolism pathway of UA biodegradation by B. paramycoides-YC02 and offer a potential avenue for the development of bacterioactive drugs against HUA and gout.High serum uric acid levels, known as hyperuricemia (HUA), are associated with an increased risk of developing gout, chronic kidney disease, cardiovascular disease, diabetes, and other metabolic syndromes. In this study, a promising bacterial strain capable of biodegrading uric acid (UA) was successfully isolated from  TherefoCurrently, the management of HUA involves pharmacological treatment and dietary intervention. Chemical drugs used for HUA treatment can be classified into three main categories: xanthine oxidase (XOD) inhibitors , uricosuric agents , and enzyme therapies (such as rasburicase and pegloticase) . Their mPhellodendri amurensis cortex, Semen coicis, Radix achyranthes root, Atractylodis rhizoma) [Astragalus membranaceus [Sanghuangporus vaninii and Inonotus hispidus [In addition, traditional Chinese medicine has been used to manage HUA and gout from a long time ago, such as Simiao Powder (rhizoma) , Compounrhizoma) , Astragaranaceus , Sanghuahispidus . Chinesehispidus . Moreovehispidus . Comparehispidus .In recent decades, an increasing number of studies have demonstrated that diet plays a crucial role in the development of HUA and gout. However, dietary interventions necessitate restricting the consumption of purine-rich foods, alcohol, and fructose. The recommended daily intake of purines in the diet is less than 400 mg in Japan. Excessive intake of purine-rich foods can lead to elevated serum UA levels , which iLactobacillus plantarum Q7 (from yak yogurt) [Limosilactobacillus fermentum JL-3 (from Jiangshui) [Bacillus had the ability to produce uricase, including B. subtillis [B. licheniformis [B. thermocatenulatus [B. cereus [According to the available research, the microbial biodegradation of UA provides new research ideas for the treatment of HUA and gout. Some studies showed that lactic acid bacteria (LAB) could biodegrade UA and had the potential to intervene in the treatment of HUA. These LAB were isolated from various fermented foods, such as  yogurt)  and Limoangshui) . But morangshui)  and suppangshui) ; many LAubtillis , B. lichniformis , B. therenulatus  and B. c. cereus , but theBaijiu cellar mud and identified as Bacillus paramycoides-YC02. The culture conditions were optimized, and both YC02 and its cell-free extract (CE) demonstrated the effective removal of UA, indicating the production of UA-biodegrading enzymes by YC02. Subsequently, the draft genome of YC02 was sequenced to find genes encoding UA biodegradation enzymes, leading to the elucidation of the biodegradation mechanism. These findings hold significant importance in the development of bacterioactive drugs targeting HUA and gout.In this study, a bacterial strain that has a better ability in UA’s biodegradation is first isolated from Baijiu cellar mud of Shandong Bandaojing Co., Ltd.  in China using UA as the sole carbon and energy source.UA with a purity of 99% was purchased from Aladdin Chemical Co. , and all other chemicals used in this study were analytical grade. A bacterial strain used in this study was isolated from 4Cl, 0.5; vitamin B complex tablets, 0.01; Na2HPO4, 0.5; KH2PO4, 0.05; MgSO4, 0.1; CaCl2, 0.01; C6H11FeNO7, 0.01; trace element solution 0.5 mL and H2O 1 L. UA at different concentrations as the sole carbon and energy source were added in the modified MSM. The Luria–Bertani (LB) medium for culture contained (g/L): NaCl, 10; Peptones, 10; Glucose, 5; Yeast powder, 5 and H2O 1 L. The initial pH of the modified MSM and LB medium was adjusted to 7.0 using 0.1 M NaOH solution. Both were sterilized in an autoclave  at 121 °C for 20 min.The modified mineral salt medium (MSM) for the isolation of bacteria contained (g/L): NHBaijiu cellar mud samples were added to 100 mL of sterilized water, stirred thoroughly and left for 30 min; then, 5 mL of supernatant was taken into a 250 mL flask that contained 45 mL of modified UA-MSM and incubated for 7 days at 38 °C with the shake rate of 200 rpm in an incubator shaker. Every 7 days, 5 mL of the cultures was subcultured to fresh modified UA-MSM with the same culture conditions each time. The concentration of UA was increased from 0.5 to 4 g/L  [Ten grams of , 4 g/L) . After 5The morphology of YC02 was observed by a microscope . The YC02 was inoculated in LB medium and incubated for 48 h at 38 °C with the shake rate of 200 rpm. Then, 5 mL cultures of YC02 were used to extract genomic DNA with a bacterial genomic DNA kit. By using the extracted bacterial genome as the template, a pair of universal primers 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492R (5′-GGTTACCTTGTTACGACTT-3′) were used to perform PCR [The culture solution of YC02 was prepared, and the bacterial precipitate was collected by the centrifugation of 10,000 rpm for 20 min at 4 °C, which was sent to Meiji Bio Co.  for sequencing the draft genome .600nm) was measured to represent the bacterial growth. In this experiment, three factors of initial pH, temperature and initial UA concentration were tested as independent variables to investigate their effects on the biodegradation ratios of UA by YC02. The initial pH of modified MSM was set at 5.0, 6.0, 7.0, 8.0 and 9.0 ; temperature at 20, 25, 30, 38 and 40 °C ; and initial concentrations of UA at 100, 200, 500, 1000 and 1500 mg/L . Every 12 h, 2 mL of cultures was taken for measuring UA concentration and OD600nm.The biodegradation experiments by YC02 were carried out in 20 mL sterilized modified MSM containing 500 mg/L UA and grown at 38 °C with the shake rate of 200 rpm for 72 h. The optical density  . The samThe monoclonal colonies of YC02 were grown on the LB agar plate  left, whBacillus paramycoides, which was identified as a new species within the B. cereus group in 2017 by Yang Liu et al. [B. paramycoides-YC02 based on phylogenetic analysis of the 16S rDNA sequence. B. paramycoides was reported to biodegrade many organics. For example, a report showed that B. paramycoides could first degrade acephate to methamidophos, and then methamidophos was degraded to some small molecules by this bacterium [B. paramycoides could utilize polyethylene as the sole carbon source [B. paramycoides could produce a variety of enzymes [B. paramycoides have been documented.According to u et al. . The stracterium . Anothern source . In addi enzymes ,48,49, s enzymes  and wast enzymes . To the The effects of different initial pH, temperature and initial concentration on the growth and the biodegradation ratios of UA by YC02 were investigated. UA biodegradation ratios exceeded 97% at initial pH values of 7.0 and 8.0, with no significant difference observed in YC02 growth a. These B. paramycoides-YC02 could completely remove 500 mg/L UA within 48 h; meanwhile, the OD600nm could reach above 1.2 and the logarithmic growth period of YC02 began at 12 h  were categorized into 21 different categories of COG a. NotablB. paramycoides-YC02 was found to be able to biodegrade UA. According to the results mentioned above, the genes and enzymes involved in the direct conversion of UA to allantoin were not found. But the flavin adenine dinucleotide (FAD)-dependent urate hydroxylaseen encoded by gene2008 was found in the GO database. Meanwhile, riboflavin kinase and flavin mononucleotide (FMN) adenylyl transferase encoded by gene1779 were found in the KEEG database. Based on these results, UA’s biodegradation pathway was identified  could decrease the serum UA levels in hyperuricemic mice [B. paramycoides-YC02 has potential promising applications in the prevention and treatment of HUA. In the following research, we also want to explore the potential therapeutic ability of B. paramycoides-fermented different Chinese herbal medicines on HUA, especially homologous medicines and foods (such as Poria cocos (Schw.) wolf, Polygonatum mill and Astragalus membranaceus).At present, fermented Chinese herbal medicines with microorganisms for the management of HUA is also a focus of our current research. Ruoyu Wang et al. found that mic mice . Recentlmic mice . BFA couB. paramycoides-YC02, an efficient bacterium for biodegrading UA, from Baijiu cellar mud. Under the optimal culture conditions , YC02 completely biodegraded an initial UA concentration of 500 mg/L within 48 h. Moreover, the CE of YC02, containing 5.38 g/L protein, successfully removed 387 mg/L of UA within 12 h. These findings clearly demonstrate YC02’s remarkable capacity for UA biodegradation. Importantly, the draft genome analysis revealed the presence of gene1779 and gene 2008, encoding riboflavin kinase, FMN adenylyl transferase and FAD-dependent urate hydroxylase, which are involved in UA biodegradation. Notably, FAD-dependent urate hydroxylase plays a crucial role in the biodegradation process by converting urate to HIU with the assistance of FAD. Subsequently, HIU spontaneously breaks down into OHCU and (S)-allantoin. These findings shed light on the metabolic pathway employed by YC02 in UA biodegradation. Additionally, our investigation unveiled numerous genes that encode enzymes responsible for the biodegradation of UA precursors and carbohydrate-active enzymes. These findings open up new avenues for future research on decreasing serum UA levels, including the biodegradation of inosine, guanosine and XOD inhibition. Not only that, we can also combine YC02 with Chinese herbal medicines to develop functional foods. We anticipate that these results will provide valuable insights into the amelioration of HUA and gout.In this study, we firstly isolated


HTLV-1 gene products highly immunogenic to humans and targets for the humoral and cellular immunity. Actually, antibodies against the HTLV-1 gag and env gene products are ubiquitously detected in HTLV-1 carriers. Furthermore, cytotoxic T lymphocytes (CTLs) against HTLV-1 Tax (TaxCTLs) are often observed in long-term survivors with ATLL treated with or without allogeneic hematopoietic stem cell transplantation. Thus, the activation of antitumor cellular immunity may play an important role among long-term survivors with ATLL, an intractable disease.Aggressive type adult T-cell leukemia/lymphoma (ATLL) caused by human T-cell lymphotropic virus type 1 (HTLV-1) infection is associated with dismal survival, even after the approval of mogamulizumab . A large number of genetic and epigenetic abnormalities as the results of the pleotropic effects of HTLV-1 Tax and HBZ reportedly are present in ATLL patients , which mIn patients with ATLL, the CD4/CD8 ratio in T lymphocytes is often inverted due to a decrease in CD4-positive T lymphocytes, including regulatory T lymphocytes (Treg), after treatment with mogamulizumab. Conversely, we have noticed that the CD4/CD8 ratio is almost always inverted in long-term surviving ATLL patients in the pre-Mogamulizumab era. Furthermore, several of these patients developed herpesvirus infections, such as herpes zoster and herpes encephalitis, while their disease was controlled by chemotherapy and subsequently exhibited CD4/CD8 ratio reversal . Herpesvin vivo. Even if the ATLL cells do not express Tax, it is possible that the large number of genetic alterations caused by Tax and HBZ could possibly lead to the expression of tumor antigens other than Tax and be captured by the host cellular immunity.Tax reportedly plays an important role in the process of ATLL development, but after ATLL development, Tax expression is not necessary for ATLL cell survival, and ATLL cells in some ATLL patients do not express Tax . HoweverTCR gene transferred T lymphocytes, and promising results have been reported for each [There have been attempts to treat patients by using Tax peptide-pulsed dendritic cell vaccine and adoptive immunotherapy by the Tax-specific for each , 11. HowIn the VAT study, for safety reasons, attenuated VZV vaccination was performed after the confirmation of the presence of anti-VZV antibodies and adequate cellular immunity as confirmed by the lymphocyte blastoid transformation test, and no adverse reaction was observed. These results may serve as a guide when considering attenuated VZV vaccination in patients with all cancer types.gag and env gene products, but these antibodies cannot eliminate HTLV-1-infected cells. TaxCTLs are also produced, but HTLV-1-infected cells escape the surveillance of TaxCTLs by controlling Tax expression and accumulate genetic abnormalities over time, eventually becoming ATLL cells. Although contracting herpes simplex or herpes zoster is unpleasant, the mechanism by which these herpesvirus infections can produce a therapeutic effect on refractory ATLL via the activation of the host’s cellular immunity is extremely interesting and worth further study.The HTLV-1 infection is recognized by the host immune system, which produces antibodies to


Cerebral cavernous malformations (CCMs) arise when capillaries within the brain enlarge abnormally, causing the blood–brain barrier (BBB) to break down. The BBB serves as a sophisticated interface that controls molecular interactions between the bloodstream and the central nervous system. The neurovascular unit (NVU) is a complex structure made up of neurons, astrocytes, endothelial cells (ECs), pericytes, microglia, and basement membranes, which work together to maintain blood–brain barrier (BBB) permeability. Within the NVU, tight junctions (TJs) and adherens junctions (AJs) between endothelial cells play a critical role in regulating the permeability of the BBB. Disruptions to these junctions can compromise the BBB, potentially leading to a hemorrhagic stroke. Understanding the molecular signaling cascades that regulate BBB permeability through EC junctions is, therefore, essential. New research has demonstrated that steroids, including estrogens (ESTs), glucocorticoids (GCs), and metabolites/derivatives of progesterone (PRGs), have multifaceted effects on blood–brain barrier (BBB) permeability by regulating the expression of tight junctions (TJs) and adherens junctions (AJs). They also have anti-inflammatory effects on blood vessels. PRGs, in particular, have been found to play a significant role in maintaining BBB integrity. PRGs act through a combination of its classic and non-classic PRG receptors (nPR/mPR), which are part of a signaling network known as the CCM signaling complex (CSC). This network couples both nPR and mPR in the CmPn/CmP pathway in endothelial cells (ECs). Cerebral cavernous malformations (CCMs), among the most common brain vascular malformations, are characterized by abnormally dilated intracranial capillaries resulting in the destruction of the blood–brain barrier (BBB), leading to hemorrhagic stroke ,2. ThreeThe BBB, the blood vessel barrier in the brain, is a complex, dynamic, and selective interface between the bloodstream and central nervous system (CNS) ,13,14. TIt is crucial to comprehend the molecular mechanisms underlying the signaling pathways that regulate and maintain blood–brain barrier permeability in both physiological and pathological states. The elucidation of the molecular mechanisms involved in the disruption of the BBB can provide insights into the effects of various factors, including abusive drugs, toxins, and pathogens. This knowledge can also be instrumental in understanding numerous neurological pathologies, such as encephalitis, meningitis, and, in particular, hemorrhagic stroke ,28. The Steroids, which are either derived from dietary sources or synthesized within the body, are defined by their distinctive four-ring carbon structure and play vital roles as hormones in various physiological functions. These compounds have been employed in numerous therapeutic applications for a broad array of human disorders ,39,40, sAt present, tissue plasminogen activator (tPA) is the most effective medication recommended for treating ischemic strokes. tPA helps dissolve blood clots, restoring blood flow to the brain. Surgery remains the only viable option for treating hemorrhagic strokes, highlighting the importance of preventing and managing strokes ,48,49,50This article provides a comprehensive review of the interplay among steroids, EC junctions, and blood vessel permeability, including the impact of inflammation on BBB integrity and the effects of angiogenic factors on BBB integrity. It also discusses the potential of steroids as future therapeutics and examines the effects of sex steroids on neovascularization and downstream angiogenic factors of PRG signaling. Furthermore, it highlights PRG’s neuroprotective effects on the BBB and the actions of PRG and its derivatives as neurosteroids. Lastly, the article explores the impact of PRG-mediated signaling on endothelial cell function and the maintenance of BBB integrity by the newly defined CmPn/CmP signaling networks.Blood vessel permeability can be influenced by various factors, such as the integrity of endothelial cell junctions, the occurrence of inflammatory events, the expression of adhesion molecules, the activation of intracellular signaling pathways, and the release of angiogenic factors. Additionally, the presence of vascular pores or fenestrations, vesicular transport, and cell–cell communications can play a role in regulating blood vessel permeability. In simple terms, the permeability of blood vessels is controlled by molecular processes that regulate the EC junction to maintain stability in the vasculature ,53. ImpaAs mentioned earlier, EC junctions mainly consist of TJs and AJs . TJs conInflammation is widely recognized as a significant contributor to the BBB dysfunction ,69. LeukVEGF plays a crucial role in regulating vascularization from embryogenesis to adulthood, including the formation of adult blood vessels ,74. In mSteroids have been found to influence the BBB through their regulation of VCAM-1 expression . ESTs haGCs are able to specifically have effects on the vasculature upon binding GRs through nitric oxide biosynthesis ,88, furtESTs belong to another group of important regulatory hormones of BBB permeability. They protect the BBB before menopause, but may increase BBB permeability with aging ,97,98. FEvaluation of female hormone contraceptives, such as PRG and its derivatives, are yet to be explored for effects on hemorrhagic stroke management and prevention. A recent report indicated that mifepristone (MIF), a widely used contraceptive and a known antagonist of PRG-nPR signaling, can inhibit PRG actions through nuclear PRG receptors (nPRs) in cells that express them. However, in cells that lack nPR expression, MIF can function as a PRG agonist on its own or in combination with PRG to activate PRG signaling through non-classic membrane PRG receptors (mPRs), leading to specific mPR-mediated PRG actions ,124,125.Hence, MIF and PRG collaborate to amplify mPR-specific PRG action in this signaling cascade . Within When damage to the endothelium takes place, EST aids in repairing the EC by reestablishing the tight and adherens junctions, which results in a decreased likelihood of hemorrhagic stroke. This finding has contributed to the utilization of tamoxifen in treating vascular conditions ,129. A sLike ESTs and GC, the effect of PRG and its metabolites/derivatives (defined as PRGs), as well as contraceptive derivatives (progestins), on BBB integrity has also been primitively explored, despite a lack of understanding that different progestins used in contraception exhibit differential off-target effects via steroid receptors . PRGs weNeovascularization is the formation of new blood vessels, which typically occurs in specific physiological processes, such as the menstrual cycle and tumorigenesis ,149. DisThe uterus vasculature is sensitive to cytokines, growth factors, and sex hormones, affecting angiogenesis and tissue growth. VEGF distribution changes during the menstrual cycle may impact angiogenesis. Endometrial stromal cells and myometrium smooth muscle cells produce VEGF, which increases in rat uterine tissue due to PRGs and ESTs ,154,155.Moreover, research has demonstrated that PRGs can increase the actions of nitric oxide (NO) in vascular ECs exclusively through membrane progesterone receptor (mPR) signaling. In a study of human umbilical vein endothelial cells (HUVECs), the role of mPRs in angiogenesis was investigated . As an mThe NVU is composed of neurons, astrocytes, ECs, pericytes, microglia, and basement membranes ,23,52,57In addition, PRG is known to regulate microglial activation, reduce the production of proinflammatory cytokines, increase the expression of antiapoptotic Bcl-2 protein, promote myelinization and myelin repair , and regThrombin has the ability to impair both endothelial cell (EC) junctions and the extracellular matrix (ECM) ,175. ThrActivated MMPs have been linked to a compromised blood–brain barrier (BBB). This is believed to be due to their ability to affect the integrity of the BBB by breaking down and restructuring the extracellular matrix that surrounds the capillaries in the brain, as well as by degrading the proteins responsible for tight junctions ,178,179.Like EST, PRG can attenuate LPS-stimulated inflammation in a dose-dependent fashion by suppressing the LPS-induced NF-κB activation. These specific anti-inflammatory effects of PRG can be inhibited by MIF, suggesting PRG-nPR as a sole anti-inflammatory signaling pathway .2+) levels by activating a specific membrane receptor called PGRMC1. This activation results in reduced nuclear accumulation of Ca2+-dependent nuclear factor of activated CD8+ T cells 1 (NFAT1), associated with T-cell development and activation. Therefore, PRG and its non-classic membrane receptor, PGRMC1, may directly suppress T-cell activation in inflammatory responses, indicating their involvement in non-classic PRG signaling [+ T helper (Th) cells, shortening the active course of some chronic autoimmune inflammatory diseases in the brain [PRG can potentially regulate intracellular free calcium  cells can be summarized with a model of CSC-modulated classic and non-classic PRG signaling . As a seIn this review, we attempted to summarize steroid signaling pathways that regulate BBB integrity through the impacts of certain steroid receptors on its key components, with special emphasis on the combined effect of PRG and CSC through the CmPn/CmP signaling network , Table 1The BBB is an intricate interface between blood flow and brain tissue with significant contribution to brain injuries, such as hemorrhagic stroke, through its disruption. Understanding the mechanisms through which various pathologies alter BBB integrity will allow for future advances in therapeutics for neurovascular injuries. Steroids, especially sex steroids (mainly ESTs and PRGs), have shown some major impacts on EC repair, neovascularization, and maintenance of BBB integrity. PRG is perhaps the most significant factor involved in maintaining vasculature and protecting neurons and the BBB. The CSC’s essential role in coupling and modulating nPR and mPR signaling was found to be integral in maintaining the BBB integrity within the CmPn/CmP signaling networks. Perturbation of this network, especially in CmP signaling in cerebral ECs, disrupts EC junction components and leads to compromised BBB. In addition to PRG, we described the role of other steroids in BBB maintenance, as well as potential roles in hemorrhagic stroke treatment.


The desired number of children is markedly higher in Sub-Saharan Africa (SSA) than in other major regions. Efforts to understand how and why these desires are generated and maintained have yielded a broad research literature. Yet there is no full picture of the range of contextual, cultural, and economic factors that support and disrupt high fertility desires.This scoping review synthesizes thirty years of research on the determinants of fertility desires in SSA to better understand what factors underlie men and women’s stated fertility desires and how they weigh the costs and benefits of having (more) children.We identified and screened 9863 studies published from 1990 to 2021 from 18 social science, demographic, and health databases. We appraised determinants of fertility desires from 258 studies that met inclusion criteria according to their roles as traditional supports or contemporary disrupters of high fertility desires.We identified 31 determinants of high fertility desires, which we organized into six overarching themes: economy and costs; marriage; the influence of others; education and status; health and mortality; and demographic predictors. For each theme, we summarize ways in which the determinants both support and disrupt high fertility desires. We find that high fertility remains desirable in many regions of sub-Saharan Africa but contemporary disrupters, such as the economic situations and increases to family planning and education, cause individuals to decrease their desired fertility with such decreases often viewed as a temporary adjustment to temporary conditions. Most included studies were quantitative, cross-sectional, and based on survey data.This review demonstrates how traditionally supportive and contemporary disruptive forces simultaneously influence fertility desires in sub-Saharan Africa. Future studies analyzing fertility desires in sub-Saharan Africa should be informed by the lived experiences of men and women in this region, with qualitative and longitudinal studies prioritized.The online version contains supplementary material available at 10.1186/s12978-023-01627-7. In sub-Saharan Africa, both men and women continue to desire large numbers of children in contrast to most regions of the world where the desired number of children is near or below replacement level (around 2.0 children per couple). We conducted a comprehensive review of the existing research on the sources and drivers of the persistently high desired fertility in sub-Saharan Africa. Based on the review, 258 studies were included in this analysis. Several factors that influence fertility desires among men and women in this population were identified. We categorized them as either traditional supports  or contemporary disrupters (factors that have more recently inhibited or discouraged high fertility desires). Although fertility desires are shaped by a wide range of factors that vary based on specific country and population, several overarching conclusions were clear. Contemporary changes in the economy and family have caused individuals to shift their fertility desires downward, and this shift is often viewed as a temporary adjustment to temporary conditions. Increased autonomy and formal schooling of women and availability of family planning can help position women to implement lower fertility desires. There is a need for more longitudinal research in this region to better understand how fertility desires may change over a person’s life, and for more qualitative studies that allow people to describe and explain their lives and fertility desires more directly in ways that allow for uncertainty and ambiguity.The online version contains supplementary material available at 10.1186/s12978-023-01627-7. Sub-Saharan Africa (SSA) is the major region where fertility rates remain high . Survey For decades, social and health scientists, among others, have investigated the drivers and motivators behind high fertility desires in SSA. Most of this research has consisted of analysis of survey data, primarily Demographic and Health Surveys (DHS). This has been complemented by a smaller number of studies employing non-survey approaches, including qualitative interviews and ethnography. The pertinent research literature is diverse methodologically, with clear variation within and across countries and regions in SSA. Hence, no small set of specific studies provides a full picture of the range of contextual, cultural, and economic factors that drive fertility desires in SSA.The goal of this scoping review is to synthesize research evidence from the last thirty years on the determinants of fertility desires in SSA. Specific research questions include: (1) What factors underlie men and women’s stated fertility desires? and (2) How do men and women weigh the costs and benefits of having (more) children? We consider how the existing literature has addressed these questions and provide a more expansive understanding of the influences of fertility desires in SSA. We also highlight gaps that future research should address. The theoretical framework guiding this review makes a basic distinction between traditional supports for high desired fertility and factors that disrupt these traditional supports, thereby prompting revised evaluations of the benefits and costs of childbearing.In the 1970s and early 1980s a prevailing view among scholars was that the high desired fertility in SSA, evident in both survey data and other types of empirical investigations, reflected the dominant social and cultural systems characterizing the region. Shaped by these systems, high fertility was rewarded, contributing to the normalization of high fertility desires and large family sizes. High fertility was perceived as important for the perpetuation of individual lineages, enlarging kinship groups and the survival of the extended family that was crucial for livelihoods (e.g. access to land) and other privileges dependent on personal connections –7. ContiAn alternative view was that the high fertility desires in SSA were not principally a function of social and cultural systems but, rather, a rational response to the continent’s specific history with colonialism . Faced wprima facie evidence that the forces of the aforementioned traditional and historical influences that shaped fertility desires in SSA over many generations are now challenged by new societal developments that have disrupted these longstanding influences.Theoretical frameworks for fertility transition that have enjoyed some success in explaining fertility declines since 1950 in other regions have proven inadequate in explaining the slow, and sometimes non-existent, progression of fertility decline in SSA , 13. WitWhat demographic and socioeconomic changes are acting to drive down desired fertility? Many of the candidates are readily recognized and have been the subject of extensive empirical investigation. These include changes in the economy, increasing importance and cost of children’s education , sharp rWith this scoping review, we aim to provide a more comprehensive understanding of the full array of influences on fertility desires in SSA. Our review of the immense literature during the past thirty years is organized around two major theoretical themes:   (1) traditional supports of high fertility desires and (2) contemporary disruptors.We conducted this scoping review guided by the framework proposed by Arskey and O'Malley  and furtWith the help of two experienced research librarians, we developed, pilot tested, and refined a search strategy pertaining to the population (men and women of reproductive age), concept (fertility desires), and context of interest (SSA). We used the search strategy to identify relevant social science, demographic, and health literature published from 1990 to 2021 in 18 databases . We searched each database using free-text keywords and controlled vocabulary specific to the database. We ran the initial search in July 2020 and updated it to include more recent literature in April 2021. We include a sample search strategy used for the PubMed database in Additional file We include studies conducted with men and/or women of reproductive age (15–49 years old). This limits the review to studies conducted among men and women for whom the topic of interest is most salient. We excluded studies that focused exclusively on special populations defined by specific health conditions that may affect their fertility intentions .We included studies if they assessed or discussed factors that affect participants’ fertility desires, excluding studies focused on family planning or contraception that did not specifically address determinants of fertility desires. Due to the wide range of disciplinary fields included, fertility desires were operationalized in several ways across studies. Table The review covers studies conducted in SSA from 1990 to 2021. The 1990 cut-off date was chosen because the DHS Program, the basis of most literature on fertility desires in SSA, was launched in Africa in 1986, with the first set of articles from the data being published in the early 1990s.We included all study types—quantitative, qualitative, and mixed methods—in the review, and incorporated a diversity of publication types including journal articles, book chapters, conference papers, dissertations, working papers, and research reports.We used the citation manager Zotero to deduplicate all search results. We then used the online systematic review software Covidence for screening and data extraction of the remaining search results. Two reviewers independently screened each search result. First all titles and abstracts were screened for relevance. Next, the full texts of all relevant results were assessed against the inclusion and exclusion criteria. We resolved disagreements at each stage through discussion until consensus was reached. When the two reviewers could not come to an agreement, a third reviewer helped resolve disagreements. This process yielded 258 included studies. Figure We extracted data from all studies that met the inclusion criteria using a standardized form developed specifically for the review. We pilot tested the form on a selection of ten studies and refined it. Two reviewers then independently extracted data from all the included studies using the finalized form. Extracted data included study characteristics, sample characteristics, operationalization of fertility desires, and study findings on fertility desires. Findings from the studies were coded and narratively synthesized according to identified determinants of fertility desires  in SSA are represented, with West Africa having the largest concentration. Figure Most pieces were published after the year 2000. Quantitative studies were overrepresented, constituting 154 of the 258 pieces. There were 64 qualitative studies and 36 mixed methods studies. Table Children have long been viewed as economic resources for families and as providing security for parents in old age in SSA. Most studies (n = 63) found that viewing children as economic resources and children providing security for their parents in old age (n = 51) increased desired fertility. Having many children provides families with additional labor, and this economic value of children is especially high in agricultural societies, rural areas, and for poorer families. Amongst these groups, having many children can help with farming and trading. In the absence of governmental social support programs for the elderly in SSA, support and care from children remains important and not easily replaceable. Thus, having children to take care of elderly parents is important, and this is particularly true for women, especially if their spouse dies –27. HaviIncreasing urbanization and changing economic conditions have altered the value of children. Almost all included studies on economy and costs (n = 60) found that recent economic changes, which have had the effect of increasing the cost of children, have decreased desired fertility. While children are still considered a source of wealth in more rural parts of SSA, they are increasingly seen as expensive burdens in urban areas where the cost of raising children and a higher overall cost of living outweighs potential economic returns. Widespread unemployment and increases in the cost of living have shaped the perceived value of children for urban residents of SSA. Among urban residents, education is perceived as improving future prospects for their children, a perception less prevalent in rural areas where farming and other agricultural occupations are still very common and require little, if any, formal schooling. Furthermore, many urban residents choose to have fewer children due to high schooling fees, so they can afford to invest more heavily in each child –33.Several studies (n = 10) found that environmental factors—natural disasters, droughts, long-term climate change—have decreased desired fertility. Several mechanisms account for this. First is food shortages, leading to a recognition that fewer children can be supported –36. SecoEconomic downturns often lead to increased migration for work and other opportunities. Migration is in turn associated with fertility desires, but this literature is limited (n = 4). Women married to successful migrants  were less likely to want to stop childbearing compared to women married to unsuccessful migrants –46. SpouSeveral pieces (n = 74) investigate fertility desires in relation to type of marital union (monogamous vs. polygynous). This collection of studies shows that polygyny both encourages higher fertility desires as well as serves as a mechanism for realizing high fertility desires. The bulk of studies (n = 47) found that polygyny was associated with higher fertility desires. Men can acquire additional wives with the purpose of achieving their desired family sizes and compositions. A small number (n = 6) found no significant difference in fertility desires between monogamous and polygynous unions. Additionally, preferences for male children may motivate entrance into polygynous unions, and high aggregate fertility desires have been found to be significantly associated with the presence and prevalence of polygyny in a community . QualitaApart from the matter of union type, several studies (n = 29) indicate that women's efforts to promote marital stability and secure spousal rights  were common reasons women wanted more children, with the caveat that too many children can also strain a marriage. A break-up of the union would place extra burden on the women for feeding, educating, and generally caring for their children. Some women felt pressure to conceive, especially after an abortion or fetal loss, to avoid marital strife . When a Several studies (n = 15) found that spousal influence increases desired number of children, with both sexes willing to defer childbearing decisions to the more pronatalist partner. With men likely to be more pronatalist and with a wife’s fertility intentions more likely to be influenced by her husband’s fertility desires than vice versa, spousal influence often plays out in gendered ways . AdditioSmaller ideal family sizes have been found among divorced/separated and widowed women compared to currently married ones , and remForty-four studies found that high rates of infant and child mortality increase desired fertility. Both men and women endorse having many children where mortality rates are high . Those wWhile we excluded studies that examined the fertility desires of HIV + individuals for reasons already described, we did include studies (n = 20) that examined how the context of the HIV/AIDS epidemic shapes fertility desires for HIV − individuals. There is a great deal of fear and stigma associated with HIV/AIDS, even for those who are not positive themselves. The sex of the participant influenced the effect of HIV/AIDS on desired fertility with women being more likely to fear HIV infection and, thus reduce desired fertility , 109. KnWomen’s perceptions of their own health also affect their fertility desires. Increased awareness of the detrimental health impacts of high parity and short birth spacing—awareness promoted by formal schooling and family planning programs—leads to reduced desired fertility , 110–114This theme encompasses the influence of other individuals or groups beside spouses on people’s fertility desires as well as the influence of social and cultural norms on fertility desires. Some studies found that larger families of origin were associated with a higher likelihood of desiring more children compared to respondents with smaller families of origin , 116; thSeveral studies (n = 22) found that desired fertility is influenced by family networks, and this effect varies according to which member of the family is the primary source of influence. For example, mothers-in-law or individuals’ own mothers may be particularly salient influences for increasing desired fertility . Young pMany studies n = 48) found that having many children can bring social status and prestige to families and communities. Large family size can symbolize and bring wealth, influence, and respect for men and women, and having many children can expand their social networks and may elevate them above their peers  found th. ObservaThree quantitative studies found no relationship between community influence and desired fertility; in these studies, community influence was operationalized as adolescent’s and women’s social interaction with their peers and/or community , 97, 123Another cultural and social norm that can influence fertility desires is gender/sex preference. In many communities, male offspring are more highly valued than female, and this preference has an effect on fertility desires, with extended childbearing in order to have male children a common practice . Most stAlmost all articles that examined the influences of a lineage or clan (n = 27) found that it increased the desired number of children. For example, in Uganda, individuals belonging to families or ethnic groups with strong clan linkages wanted an average of 10.5 children, in contrast to those lacking such strong linkages who wanted 4.9 children . This inFindings from studies that examined the effect of ethnicity on fertility desires varied widely due to regional variation in included studies. Generally, ethnic groups within which large numbers of women preferred to have no more children are concentrated in East and Southern Africa , 133. InSeveral studies (n = 55) found that the association between religion and desired fertility was influenced by specific religious affiliation and sex. In general, for both men and women, being Muslim is associated with higher fertility desires compared to other religious groups, although effects are sometimes stronger for men , 136. InStudies that explicitly measured the effects of colonialism, modernization and/or Western influence were uncommon. In some focus group discussions, participants mentioned that there was an increase in individualism and copying of Western behavior and thus traditional kin obligations may no longer be strong enough to sustain the demand for high fertility, especially as participants perceived that extended family support systems are eroding . Small-fA very common finding (n = 90) is a negative relationship between formal schooling and desired fertility. Two studies found that higher education was associated with high fertility , 141. InHigher education is a mechanism for women gaining status outside of the home and, more concretely, acquiring their own source of income, which in turn can affect their fertility desires. Findings regarding the effects of the status of women on fertility desires were somewhat disparate due to a lack of standardization in how the status of women was measured, including autonomy, empowerment, education, household profit contribution, and bargaining power. Several studies (n = 21) found that fertility desires fell with improved status. For example, women who contributed more than 50% to household expenditures are less likely to desire more than four children compared to women who contribute less than 50% . On the Increased education also increases knowledge of and access to family planning. Family planning and contraception use, or positive attitudes toward use, were associated with decreased desired fertility in 32 studies. Women who discussed family planning with their husbands were more likely to limit childbearing than those who never discussed it , 144. CoSeveral studies (n = 8) found that general exposure to mass media decreased desired fertility. Watching TV and listening to the radio have been associated with decreased fertility desires and higher likelihood of wanting to stop childbearing , 150. MaA large number of included studies examined the relationship between age, parity, sex, and fertility desires. Age and parity are clearly related to each other , hence both determinants influence fertility desires in similar ways. For both of these predictors, the direction of the relationship depends on how fertility desires are operationalized. For example, in 39 studies, increasing age decreased desired number of children, with women aged 30 and above more likely than younger women to desire smaller numbers of children. On the other hand, increasing age increases ideal family size (n = 35) primarily due to post-hoc rationalization of existing children. Similarly, as parity increases (n = 47), desired fertility decreases, and the odds of wanting no more children increases with the number of living children. Women with higher numbers of living children are less likely to desire additional children compared to women with small numbers of children. As parity increases, ideal family size also increases (n = 14) due to post-hoc rationalization of past births as desired, regardless of whether they were desired at the time of the pregnancy. Seven studies found that age had no effect on desired number of children, but these studies either examined only adolescents (aged 15–19) or had small sample sizes.Other findings related to age demonstrate that the association between age and fertility desires is more nuanced and can be influenced by current economic conditions and continued modernization. Young women in particular may be under more serious pressure than previous generations in light of harsh economic realities that would imply higher opportunity costs for their time and result in lower desired numbers of children .There were consistent sex differences in desired family size (n = 43). Men were found to be more pronatalist and in favor of larger families compared to women, and this is in part due to the different roles that men and women tend to play within the home and in communities. Men often want large families because children can help with labor, while women often want smaller families because they are the primary caregivers and are more aware of the amount of work it takes to raise children .Socioeconomic status (SES) and employment also affect fertility desires. Thirty-three studies indicated that as income and wealth increase, the desire to stop childbearing increases. Women in higher wealth quintiles are more likely to desire limiting childbearing than women in poorer wealth quintiles. A smaller number of studies found that SES both increased and decreased desired fertility when temporal increases in income or wealth were taken into account , 98, 99.Regarding employment, women who work and earn income tend to want fewer children than women who do not work. Most studies (n = 31) found that the relationship between employment and desired fertility was influenced by the type of employment, including whether it was characterized as formal versus informal or agricultural versus non-agricultural. Increases in the percentage of women working in formal employment reduced preferred family size, and women’s formal employment is consistently associated with decreased desired fertility . Men’s eFertility desires in SSA have been intensely studied since the 1960s by social science and health scholars employing diverse methodologies and approaches. Collectively, this research literature has endeavored to understand how and why high fertility desires are generated and maintained in SSA. Our review of this literature shows that fertility desires are shaped by a myriad of factors, with results for any factor varying according to methodological approach and the country/population under examination. This produces some complexity, and even some contradictions, in the conclusions that can be drawn from the entire body of included studies. This feature notwithstanding, there are clear overarching patterns found in the supports and disrupters of high fertility desires. While contemporary changes in economic and family structures have influenced people’s desired fertility, causing them to shift their ideal family size downward, this is often viewed as a temporary adjustment to temporary conditions. The traditional and historical supports for high fertility, which have created strong ideological and cultural pronatalist values, have not disappeared entirely but, rather, have become more difficult to implement.There is a great deal of evidence that increased autonomy of women, increased formal schooling of women, along with improved SES decreases fertility desires and positions women to more effectively implement their low[er] desires , 157. ThWe must acknowledge that improvements in the quality of life in SSA may result initially in increases in fertility, as improved conditions mean that individuals can afford and care for the additional children they continue to desire. There is evidence that such increases in desired fertility are temporary, and fertility desires (and fertility rates) decline as positive socioeconomic conditions continue and become normalized. This evidence is limited; how fertility desires and fertility rates in SSA respond to improvements in the quality of life is an important topic for continued research, especially using longitudinal designs.why. Mixed methods approaches and increased in-depth qualitative inquiry to supplement existing quantitative survey data collection efforts will greatly contribute to knowledge production on fertility desires in the region and, importantly, provide residents of SSA the opportunity to tell their own stories and explain their lives directly. This is particularly relevant when trying to understand and/or measure uncertainty related to fertility desires, which has been difficult to achieve with existing quantitative data.We have identified several gaps in existing knowledge on fertility desires in SSA several of which can be attributed to methodological approaches. In our review, we find that 95% of included studies are cross-sectional. Longitudinal studies in the region are necessary to supplement existing cross-sectional data. We believe there should be increased, long-term funding that is necessary to undertake these kinds of projects. Additionally, it is often the case that quantitative or qualitative data alone provide a partial story. Quantitative studies predominate in our review, and many of these studies point to the need for their statistical findings to be supplemented with qualitative inquiry to better understand the mechanisms, contextualize relationships, and, ultimately, to understand Another gap in existing research on fertility desires in SSA is the limited inclusion of men. In a patriarchal society where men are often seen as authoritative, knowledge of the ways in which ideologies and norms regarding masculinity impact women’s fertility desires and outcomes is particularly important. While the inclusion of men has grown over time, and many studies do include men or even focus on men alone, there is still much to be understood about the benefits of high fertility for men and their desires relative to their spouses. Studies show that spousal communication can be an important influence on fertility desires and family planning use, but how exactly spouses communicate with each other and how this influence works is not well understood , 159. FuWe note several limitations of this review. First, we did not specifically search for grey literature. However, we searched several databases that include reports, white papers, dissertations, and other types of publications beyond journal articles, as well as the personal collection of the principal investigator which includes several decades of research on fertility thus allowing us to capture some grey literature. Although we used a wide array of search terms to capture concepts of fertility and family size desires in SSA, some articles may not have been identified. Second, due to the limited language skills of the authors, we have only included studies published in English. This resulted in the exclusion of 46 citations published in French, the official language in several SSA countries. This could have led to the exclusion of work from authors native to the region and could explain why several countries with high fertility rates  are represented by few studies. However, almost half of the 46 excluded citations were either DHS reports or duplicates of items included in English and would have therefore been excluded for other reasons. Third, due to the COVID-19 pandemic and resulting library closures and lending restrictions, we were unable to obtain the full texts of several studies which may have been relevant for this review.Based on our review of 258 articles, we conclude that traditional and historical supports for high fertility continue to influence fertility desires in SSA up to the present. Concurrently, there are important contemporary disrupters of those supports that are depressing fertility desires in the region. These disrupters could be viewed as positive  and negative  in nature. A striking conclusion of this review is that declines in desired fertility are  often a response to poor structural conditions; it follows that when those conditions improve, fertility desires and rates may respond positively, because high fertility remains highly valued in many contexts. Given the limitations of existing research and calls for methodological diversity, we believe that increased qualitative inquiry to supplement existing survey research and more longitudinal research in this area is needed. We call for future research that  centers the experiences and desires of SSA men and women without implying Western ideals or understandings of fertility and its benefits and consequences.Additional file 1: Appendix A Table S1. Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) Checklist. Appendix B Table S2. Sample Search Strategy in PubMed. Appendix C Table S3. Overview of variables’ relationship to fertility desires found in studies.


Increasing survival rates after hematopoietic stem cell transplantation (HSCT) in childhood should put focus on improving the quality of life as adults. An essential aspect is fertility and its preservation. In order to take advantage of the possibility of fertility preservation, fertility counseling should be provided to patients and their parents prior to gonadotoxic therapies.The aim of this survey was to analyze the impact of fertility counseling in pediatric stem cell transplantation in patients and their parents using questionnaires designed for the study questions. Fifty-one parents and 7 adolescent patients were interviewed between February 2019 and October 2021 about the counseling, their perceptions of fertility issues, and the nature of decision- making concerning fertility preservation. The study included patients with malignant  and nonmalignant diseases  who received counseling on fertility preservation before HSCT based on an in-house standard and analysed the impact for both groups.Two-thirds of the study participants were concerned about having children and grandchildren respectively; for half of all respondents, the topic of fertility and fertility preservation proved to be hopeful. Forty percent of the study participants were burdened by the risk of possible fertility limitations after HSCT. Concerns about fertility was particularly significant for parents whose children were advised to undergo fertility preservation. Parents of children <12 years found deciding on appropriate measures more difficult. Parents with children >7 years involved their children in the decision. All study participants agreed that fertility counseling had not negatively affected the parent-child relationship. More than 90% of all study participants were in favor of addressing fertility, its potential limitations and fertility preservation measures before HSCT. There was no significant difference between the malignant and the non-malignant cohort in all study questions.Overall, the standardized fertility counseling provided in our center of pediatric stem cell transplantation resulted in high satisfaction among patients and their parents. Multiple counseling on infertility risk, including the younger patients in the decision-making and further options after gonadotoxic therapy may increase the satisfaction of the counseled patients and their parents. Fertility is one of the essential capabilities of human beings both for the individual and for the continuity of society. Negatively influencing factors include vital therapies with a broad spectrum of side effects such as chemotherapies and radiation. Hematopoietic stem cell transplantation (HSCT) has a particularly high risk of fertility impairment as a therapy-related complication, at 75% .In the treatment of children and adolescents, fertility and its preservation prior to gonadotoxic therapy is an issue of increasing relevance. About 80% survived 30 years after diagnosis of childhood oncologic disease . SimilarIn our experience all information that parents receive during the pre-transplant interview, the risk of infertility appears to be one of the most difficult side effects to realise. Infertility means a disruption of life continuation because there will not be a next generation. Therefore, physicians have to offer various possibilities for FP that are adjusted to special needs and medical conditions of the individual patient and available medical resources. Therefore, a multidisciplinary counselling session, access to fertility preserving measures and an opportunity for their own family planning, is mandatory.Fertility counseling before chemotherapy and other gonadotoxic treatments lowers the level of fertility-related anxiety in patients and is associated with better mental health , 6. ThusThe establishment of a uniform educational session on fertility issues after HSCT as well as counseling on possible fertility-preserving measures should be understood as a self-evident and obligatory part of the pre-transplant interview. A recent study shows that despite recommendations for fertility counseling overall only 39% of patients received counseling prior to HSCT . A similCurrently, fertility counseling and the implementation of fertility-preserving measures vary within Europe depending on national recommendations, technical expertise, local circumstances, and cost coverage by the health care system –12.The questionable cost recovery for counseling and associated low staffing are reasons why not all pediatric departments handling gonadotoxic therapies have established fertility counseling to date. Different to some other countries there is no general coverage by the health insurance for fertility preserving measures.The aim of the conducted study is a survey on the quality and impact of fertility counseling in pediatric HSCT by interviewing parents and patients >12 years about their handling of fertility issues, their decision-making, as well as an assessment of fertility counseling, both, for HSCT indication for non-malignant and a malignant diseases in patients.In this monocenter, noninterventional study we included patients and parents who received fertility counseling at our center. This prospective study included all pediatric patients who received their first HSCT for malignant or non-malignant disease from February 2019 to October 2021. Attendance at previous fertility counseling appointments prior to first HSCT was not cause for exclusion. Patients who had already undergone multiple transplants and patients and their parents who could not answer the German questionnaires without a translator due to a language barrier were excluded from the study though off course they received a fertility counseling translated in mother-tongue as well.In children <12 years, the parents were invited to participate in the survey and in adolescents 12–18 years, the patient and parents were invited to participate.As a standard procedure, each patient (age-dependent) and his or her guardian received an individual 60-min counseling session by a pediatric oncologistof the HSCT team prior to HSCT. An in-house standard operating procedure (SOP) for fertility counseling implemented at our institution was used to facilitate and standardize counseling. This procedure was published in 2022 . RecommeThe counseling sessions included the individual risk assessment of infertility, description of the recommended procedures, and individualized and rational decision- making with the patient and his or her guardian. Patients who were initially counseled in the oncology department and required HSCT during the course of therapy were counseled again.The coded and anonymized questionnaire was administered to the parents and patients >12 years within the first 2 weeks of the inpatient stay in the context of the HSCT. This was done by an independent person to avoid that study participants answer in a socially desirable way.Approval for the study was granted by the Ethics Committee of the University Hospital Frankfurt am Main (business number 86/18). The study was conducted according to the principles of Declaration of Helsinki.Two versions of questionnaires were developed, one for parents and the other for patients >12 years. The study focused on three main questions: (1) Perception of fertility issues prior to HSCT, (2) Decision-making about fertility preserving measures, (3) Evaluation of the fertility counseling. For this purpose, the questionnaire had four parts. Part A contains questions about sociodemographic data; part B covered medical history and fertility counseling in the past. Some questions were directed at patients and their guardians who received fertility counseling prior to the current counseling as part of a previous therapy. This group was treated as a subpopulation. Part C addressed the perception of fertility in the context of treatment, the patient's desire to have children and the parent-child relationship. Part D included the decision-making for or against fertility-preserving measures. Last, Part E assessed the individual fertility counseling. The questionnaire included 54 and 53 questions in the version for parents and for patients >12 years, respectively.To improve the questionnaires, a pilot study had been conducted in the department of oncology as part of a partner study including 12 adolescents and parents of children 0–18 years as well as a number of healthy adolescents  to improve understanding and age-appropriate vocabulary of the questionnaire.The evaluation of the questionnaires and the data analysis were performed with IBM® SPSS® Statistics, version 28.0.0.0 and R . Data from parents and adolescents were analyzed separately. Descriptive statistics for categorical data are presented as absolute frequencies and percentages. For metric data, the mean, median, dispersion measures, and interquartile ranges were calculated and minimum and maximum were reported.U test was applied. Fischer's exact test was applied to test the independence of 2 nonmetric-scaled variables. An undirected relationship between 2 ordinal-scaled variables was tested by the Kendall-τ correlation coefficient and Spearman-rho correlation. To test the influence of an independent variable on a nominally scaled categorical variable, logistic regression analysis was used. A two-sided significance level of α = 0.05 was chosen for all statistical tests mentioned.To compare two populations that differed in the expression of an independent variable, the nonparametric Mann–Whitney A total of 92 participants were enrolled in the study. Seventy-six percent of the patient cohort were German, 6% Turkish, 6% Syrian, 4% Afghan, and 8% other nationalities. Forty-three percent reported being Christians, 31% Muslims, 6% followers of other religion, 20% did not state a religious affiliation.Forty-nine percent of the patient cohort had malignant disease, 51% a non-malignant disease. In the cohort of the no-malignant group there were 20 patients <12 years and 6 >12 years, in the malignant group 19 patients <12 years, 6 >12 years respectively.Overall, 90.2% of parents were in favor of addressing fertility and fertility preservation before HSCT. n = 30, distribution in non-malignant/malignant cohort is depicted in For 82.6% of all parents (n = 30). Further reasons were young age of the patient (42.9%), experimental character of the procedure (42.9%), and fertility was not currently a relevant issue (28.6%). Treatment delay and religious reasons were named in one case respectively . There was no significant difference between the malignant and the non-malignant cohort.In 18 of the 30 families (60%) decision-making included the child, including 2 families who decided explicitly at the child's request. Eleven of 30 parents (37.9%) found it difficult to decide. Decision-making without involving the child proved to be independent of sociodemographic factors such as gender, religion, and nationality, as well as independent of the malignancy of the child's disease. The age of the child, on the other hand, correlated strongly with parental decision making. For children <7 years of age, the decision regarding fertility-preserving measures was usually made by parents alone, and children >7 years were included in the decision-making process , 60.8% indicated that having their own children was important to their children as well . Fertility had become important to more than half of the patients due to the confrontation with this topic in the context of the disease . 39.2% of parents felt burdened by the issue of fertility , 47.1% of parents saw fertility preservation as a source of hope . In 68.6%, concerns and in 78.7%, doubts about HSCT as a therapy were not increased by fertility counseling. In 21.4% of parents concerns about HSCT were heightened, almost exclusively parents of children with non-malignant diseases . However, this did not affect opinion about the necessity of HSCT in 86.3%. Fifty percent of parents considered fertility loss important among other side effects . According to the parents, the topic of fertility had not negatively influenced the parent-child relationship. The own relationship with the child was perceived as strengthened by 11.8% of the interviewed parents.Having their own children was important to 68.6% of parents of the total study population (Forty-three of 49 families (87.8%) were satisfied with the counseling and recommended measures. 3 were rather not satisfied and another 3 could not assess their satisfaction.For 44 of 49 parents, the counseling was understandable for themselves and their child, the duration was judged sufficient by 42 of 47 parents, 87.5% were able to ask their questions during counseling. Six parents who were unable to ask their questions named emotional overload (3 parents), the presence of the child (1 parent) and unspecific issues (2 parents). 17.8%, would have preferred counseling without a child.61.7% had no suggestion for improvement of pre-HSCT fertility counseling. The most common suggestion for improvement (7 parents) was proving written information material, more appointments (5 parents), more time for counseling (4 parents) and more time on reflection (5 parents).p = 0.231), satisfaction with interventions/counseling (p = 0.920), nor on importance of fertility as a source of hope (p = 0.375) or the weighting of fertility loss among other side effects of HSCT (p = 0.846). Within the whole study cohort here were no significant differences between parents of male and female patients on any variable.There were no significant differences between parents of patients with malignant and non-malignant disease on any variable, neither on the topic of burden of fertility (p = 0.022).The correlation of the parents' desire to have children and the assessment of the child's desire to have children showed a significant correlation  who received multiple fertility counseling, if the indication for HSCT arose in the course of therapy. Within this subgroup fertility-preserving measures were recommended in 13 cases . 68.4% of parents indicated that they had become more involved with the topic since the first consultation. Slightly more than half had talked to their own child about fertility since then, while 42.1% had not. The majority of respondents found it easier to understand the content of the survey during the second consultation. The second appointment was easier for the majority and 73.2% of the subpopulation were better able to follow and understand the 2nd consultation.Opinion about fertility had not changed for 68.4% since first consultation, though familiarity with the counseling did not facilitate the decision-making for 72.6%.p = 0.018) (p = 0.055). There was no significant difference in the other variables tested, burden of addressing fertility, fertility loss as an important side effect of HSCT, and decision severity.For the subpopulation of multiple counseled families, there was a significant difference compared to the rest of the study population. Parents who had a history of fertility counseling were found to be more satisfied with the counseling and interventions than parents who were counseled for the first time in the context of HSCT (= 0.018) . FurtherThere were only 7 questionnaires of adolescents available, thus no statistical analysis was possible. All 7 patients were advised to seek fertility counseling by the oncology team or the HSCT team.Four out of 7 patients had thought about their family planning before the consultation. 4 adolescents had concerns about their fertility because of HSCT.Having their own children was important to 4 patients, 5 assumed that having grandchildren was important to their parents.For 5 out of 7 adolescents, the topic of fertility had become more important at this stage of life due to the disease. For 5 patients, their fertility and fertility preservation was a hopeful factor during the disease phase. Four adolescents felt that the possible loss of fertility was important, in addition to other side effects of HSCT, 4 felt burdened by the issue of fertility. For 2 adolescents, concerns about HSCT had increased because of the fertility issue. Four adolescents found it easy to talk to confidants about the fertility issue. All patients denied that the topic of fertility affected their parent-child relationship. Six of 7 adolescents were in favor of fertility counseling.Six of 7 patients were advised on fertility preservation measures, all opted for fertility-preserving measures when possible. The decision was made jointly with the parents by 4 patients. Three patients made the decision at their own request. One patient expressed that he made the decision at the request of his parents.Six of 7 patients were satisfied with the counseling and felt that the counseling was understandable.Fertility counseling prior to gonadotoxic treatments is highly relevant due to evolving therapies and improved survival rates. The current Society of Pediatric Oncology guideline mentions fertility preservation counseling, yet, contrary to adult medicine, there is no precise guideline for its implementation , 14. A sWith this study we sought to analyze the meaning of the counseling for the adolescent patients and their parents and how the counseling is perceived. To our knowledge this is the first study performed in this immediate setting.Considering the circumstances of just being admitted and starting therapy for HSCT and possibly the impression of the first therapy side effects such as infection or mucositis, which hinder or make participation impossible, our survey return rate is high and our study population represents the spectrum of pediatric HSCT very well though we had to exclude patients and families without secure German language skills due to the German questionnaire. In another setting it would be worthwhile to look for inter-cultural differences concerning the topic of fertility.For half of the study participants, fertility with its potential limitations after HSCT represented a burden. The existence and extent of this burden in cancer patients is also shown in a variety of other studies –19. In oMost of all respondents were satisfied with the counseling at our center, its comprehensibility and the recommended measures. Those who received counseling more than once had a significantly higher level of satisfaction. Sociodemographic differences and the type of disease of the child had no influence on the satisfaction of the parents. Similar results were found by von Wolff et al. in a survey regarding fertility concerns and counselling performance in an adult women cohort . The mosParents of adolescent patients found it easier to decide on fertility preservation measures than parents of young children. Depending on the age of the children, the decision is usually made by the parents alone and place a great burden on them. Adolescents wish to be informed about the possible loss of fertility and want to be part in treatment-related decisions . As a coIn contrast to other studies the present work found no significant difference in distress concerning decision-making between parents of male and female patients . Others The patient group with malignant vs. non-malignant diseases was differed in some aspects. Although there were no differences in burden, importance of fertility, decision severity, and satisfaction with counseling and interventions, there are differences concerning the pretreatment with potential gonadotoxic agents , risk ofParents counseled multiple times were shown to be more satisfied with both the counseling and the interventions even though it did not facilitate decision-making. A deeper understanding and good knowledge of medical interventions has been shown to lead to greater satisfaction and have a positive effect on mental health , 27.Satisfaction is also related to one's active participation in counseling and decision-making concerning the procedures and leads to less doubt about the decision and thus contributes to higher quality of life . Less doSatisfaction with one's decision is highest approximately one month after counseling . NeverthThis is a single center study with only a small number of cases. The multinational composition of the patient population limited the number of patients who could be recruited for the German questionnaire due to the language barrier. The lower response rate among adolescents can be explained in part by the difficult circumstances surrounding the survey. Parents of severely ill patients are less likely to participate in surveys. In this view the setting of our study as to be critically reconsidered and weighed against the logistical advantages of a compact setting on the HSCT ward.Thus, the survey results might only be a snapshot in the early phase of HSCT and might change with time. Poor or worsening health condition of the patients led to drop out of the study and in others may have influenced the response to the questionnaire and resulted in a lower response rate. Due to the very specific question, no validated questionnaire could be used. Larger and multicenter study trials are necessary to confirm our results especially as we had to rely on the German speaking study population only.It is desirable that every clinic treating children and adolescents with gonadotoxic therapies should also apply an SOP for counseling in clinical practice , 10. ThiFertility counseling proves to be important to patients and families and results favor multiple counseling and the involvement in the decision-making process for pediatric patients even of young age, both for patients with malignant as well as non-malignant diseases and their parents.


Teeth are unique and multifaceted tissues that are necessary for routine functions, such as crushing food and constructing articulated sounds, as well as for esthetic symbols. Nociceptive responses to external stimuli in the dentin have been detected in the dental pulp and neuron complex . SurrounHowever, an imbalance between these hard and soft tissues leads to pathological findings in oral tissues, including teeth, thereby resulting in dysfunctional physiological processes. To prevent this phenomenon, intracellular signaling control, intercellular communication, genetic regulation, and functional regenerative medicine based on tissue engineering have been considered . In a prLyu et al. studied apical periodontitis caused by bacterial infections as well as reviewed recent studies on it, focusing on the regulatory network of mesenchymal stem cells (MSCs) . Apical i) levels via Gαs protein-coupled CGRP receptor activation in odontoblasts [i response from the odontoblasts approximating the stimulated TG neurons was measured using an odontoblast–neuron co-culture system. The authors revealed the functional expression of Gαs protein-coupled CGRP receptors in odontoblasts. CGRP receptor activation increases [cAMP]i levels by the action of adenylyl cyclase. The mechanical stimulation of small-sized CGRP-positive but neurofilament heavy chain-negative TG neurons increases [cAMP]i levels in odontoblasts localized near the stimulated neurons.The functions and properties of cells are regulated by niche microenvironments. The dental pulp is a soft tissue of a cranial neural crest-derived mesenchymal origin enclosed by a rigid mineralized dentin, which ultimately resides in a low-compliance environment . When antoblasts . The auti levels in odontoblasts by activating CGRP receptors and decreased dentin mineralization. Thus, the CGRP–CGRP receptor axis plays a critical role in regulating dentinogenesis via intercellular communication. These results may provide functional evidence for axon reflexes mediated by the CGRP–CGRP receptor axis in the dental pulp tal pulp .s, Gαi, Gαq, and Gα12/13. Many receptor conformations lead to various highly specialized downstream signaling cascades. GPCRs induce two major signaling pathways, namely cAMP and phosphatidylinositol signaling. Gαs and Gαi regulate the cAMP-generating enzyme adenylyl cyclase. Gαs activates adenylyl cyclase, whereas Gαi inhibits adenylyl cyclase, thus increasing or decreasing [cAMP]i levels. Diffusible intracellular secondary messenger systems can stimulate or inhibit downstream signaling, thereby exerting further biological effects [The adenylyl cyclase signaling pathway, regulated by G-protein-coupled receptors (GPCRs), induces intracellular cAMP signaling in various cells ,13. G pr effects .s, prostaglandin I2 (IP), 5-hydroxytryptamine 5-HT4 (5-HT4), dopamine D1 (D1), adenosine A2A (A2A), and vasoactive intestinal polypeptide (VIP) receptors, immunoreactivity was observed in human odontoblasts [2+, the application of IP, 5-HT4, D1, A2A, and VIP receptor agonists increased intracellular cAMP levels. This increase was inhibited by using an adenylyl cyclase inhibitor and the abovementioned receptor antagonists in a dose-dependent manner. These results suggest that odontoblasts functionally express the Gs-protein-coupled IP, 5-HT4, D1, A2A, and VIP receptors [Kitayama et al. reported that in the presence of plasma membrane Gαtoblasts . Furthereceptors .Primary cilia are microtubule-based organelles that serve as hubs for transducing various developmental signaling pathways, including Hedgehog, Wnt, fibroblast growth factor, and platelet-derived growth factor signaling . CiliaryCollagen type I alpha 1 (Col1A1) and dentin matrix protein 1 (DMP-1) are vital matrix molecules in biomineralization ,23,24,25Collagen-based materials such as membranes, sponges, matrices, hydrogels, and composite scaffolds have been extensively used in in vivo studies to facilitate bone tissue regeneration. Takayama et al. summarized growth factor (GF) delivery systems that use collagen membranes (CMs) for bone tissue regeneration . BiomateIn this Special Issue, we discussed mineralization systems based on intracellular signaling and tissue engineering using cells, GFs, and scaffolds. We further discussed their potential applications for understanding cranial development and disease models. In the first and second editions, we additionally need to explore the underlying mechanisms of these concepts in vivo, especially in large animal models.