TAp73 knockout mice show morphological and functional nervous system defects associated with loss of p75 neurotrophin receptor.

Total and N-terminal isoform selective p73 knockout mice show a variety of central nervous system defects. Here we show that TAp73 is a transcriptional activator of p75 neurotrophin receptor (p75(NTR)) and that p75(NTR) mRNA and protein levels are strongly reduced in the central and peripheral nervous systems of p73 knockout mice. In parallel, primary cortical neurons from p73 knockout mice showed a reduction in neurite outgrowth and in nerve growth factor-mediated neuronal differentiation, together with reduced miniature excitatory postsynaptic current frequencies and behavioral defects. p73 null mice also have impairments in the peripheral nervous system with reduced thermal sensitivity, axon number, and myelin thickness. At least some of these morphological and functional impairments in p73 null cells can be rescued by p75(NTR) re-expression. Together, these data demonstrate that loss of p75(NTR) contributes to the neurological phenotype of p73 knockout mice.

Management of patients with endometriosis and infertility: laparoscopic treatment and spontaneous pregnancy rate.

OBJECTIVE: To evaluate factors involved in spontaneous pregnancy rate after surgery for endometriosis in patients with endometriosis and infertility. METHODS: This retrospective study spanned from 2014 to 2020 and included a follow-up period of two years of patients with endometriosis-related infertility who underwent laparoscopic surgery. Women aged 25 to 43 years with patent tubes, no/mild male factor and no other infertility factors were selected and grouped according to fertility management as follows: patients immediately prescribed ART (16.5%, ART-p); patients who chose not to undergo ART (83.5%) and achieved spontaneous pregnancy (71.8% SP-p); and patients who first chose not to undergo ART but had it subsequently (28.2%, NSP-p). RESULTS: A total of 200 patients were analyzed. Of the 167 patients who waited for spontaneous pregnancy, 71.8% achieved it. We observed a tendency of higher endometriosis ASRM scores in the ART-p group compared with patients who waited for spontaneous pregnancy, and lower scores in individuals that achieved spontaneous pregnancy. When we looked at how long it took to achieve pregnancy, we found that individuals in the SP-p group achieved pregnancy in 5.7 months, while subjects in the NSP-p group took 1.8 times longer than their peers in the SP-p group (p<0.001). However, once prescribed ART, the individuals in the NSP-p group achieved pregnancy within a similar time when compared with subjects in the SP-p group. In order to identify individuals that might benefit from ART early on, we performed a multivariable analysis and developed a decision tree (81.3% accuracy and 53.3% sensitivity). CONCLUSIONS: The present results indicated that, after surgery, the majority of patients achieved spontaneous pregnancy. The decision tree proposed in this study allows the early identification of patients who might require ART, thus decreasing the time between surgery and pregnancy and improving overall outcomes.

Earthquake-enhanced dissolved carbon cycles in ultra-deep ocean sediments.

Hadal trenches are unique geological and ecological systems located along subduction zones. Earthquake-triggered turbidites act as efficient transport pathways of organic carbon (OC), yet remineralization and transformation of OC in these systems are not comprehensively understood. Here we measure concentrations and stable- and radiocarbon isotope signatures of dissolved organic and inorganic carbon (DOC, DIC) in the subsurface sediment interstitial water along the Japan Trench axis collected during the IODP Expedition 386. We find accumulation and aging of DOC and DIC in the subsurface sediments, which we interpret as enhanced production of labile dissolved carbon owing to earthquake-triggered turbidites, which supports intensive microbial methanogenesis in the trench sediments. The residual dissolved carbon accumulates in deep subsurface sediments and may continue to fuel the deep biosphere. Tectonic events can therefore enhance carbon accumulation and stimulate carbon transformation in plate convergent trench systems, which may accelerate carbon export into the subduction zones.

Sperm DNA fragmentation and male age: results of in vitro fertilization treatments.

OBJECTIVE: This study aimed to assess the effects of sperm DNA fragmentation in parents belonging to different age groups. The couples included in the study comprised normozoospermic men and infertile women undergoing conventional IVF. METHODS: The results obtained from 163 conventional IVF cycles were analyzed retrospectively. The couples enrolled in the study included women aged between 30 and 37 years. Sperm DNA fragmentation was studied using the TUNEL assay. The patients were split into four groups based on male age and sperm DNA fragmentation, as follows: Group 1: ≤39 years and TUNEL assay ≤20%; Group 2: ≤39 years and TUNEL assay >20%; Group 3: ≥40 years and TUNEL assay ≤20%; and Group 4: ≥40 years and TUNEL assay >20%. RESULTS: No significant differences were found in semen parameters or fertilization rates between groups. Groups with <20% sperm DNA fragmentation showed significant differences in other parameters, including higher blastocyst formation rate (Group 1: 63% and Group 3: 60% vs. Group 2: 43% and Group 4: 41%, p<0.05) and higher expanded blastocyst formation rate (Group 1: 42% and Group 3: 40% vs. Group 2: 21% and Group 4: 18%, p<0.05). Miscarriage rate was significantly higher in Group 4 (42% and 46% vs. 5%, 25% and 5% in Groups 1, 2 and 3, respectively, p<0.05). CONCLUSIONS: Our results showed lower blastocyst formation rates from IVF when males had high levels of sperm DNA fragmentation. Higher miscarriage rates were also observed in couples with males aged 40+ years. These results reinforce the need to inform couples with male partners aged 40+ years about the potential risks inherent to fertility treatment.

Amyloid Β-Peptide Increases Mitochondria-Endoplasmic Reticulum Contact Altering Mitochondrial Function and Autophagosome Formation in Alzheimer's Disease-Related Models.

Recent findings have shown that the connectivity and crosstalk between mitochondria and the endoplasmic reticulum (ER) at mitochondria-ER contact sites (MERCS) are altered in Alzheimer's disease (AD) and in AD-related models. MERCS have been related to the initial steps of autophagosome formation as well as regulation of mitochondrial function. Here, the interplay between MERCS, mitochondria ultrastructure and function and autophagy were evaluated in different AD animal models with increased levels of Aß as well as in primary neurons derived from these animals. We start by showing that the levels of Mitofusin 1, Mitofusin 2 and mitochondrial import receptor subunit TOM70 are decreased in post-mortem brain tissue derived from familial AD. We also show that Aß increases the juxtaposition between ER and mitochondria both in adult brain of different AD mouse models as well as in primary cultures derived from these animals. In addition, the connectivity between ER and mitochondria are also increased in wild-type neurons exposed to Aß. This alteration in MERCS affects autophagosome formation, mitochondrial function and ATP formation during starvation. Interestingly, the increment in ER-mitochondria connectivity occurs simultaneously with an increase in mitochondrial activity and is followed by upregulation of autophagosome formation in a clear chronological sequence of events. In summary, we report that Aß can affect cell homeostasis by modulating MERCS and, consequently, altering mitochondrial activity and autophagosome formation. Our data suggests that MERCS is a potential target for drug discovery in AD.

Optimizing the number of embryos to transfer on day 5: two should be the limit.

OBJECTIVE: To define the appropriate number of embryos to be transferred at day 5. METHODS: Retrospective analysis of 784 consecutive fresh day-5 embryo transfers performed between 2007 and 2015, divided in three groups: Group A (N = 219): received the only 2 embryos that reached a transferable stage; Group B (N = 357): received 2 selected embryos among several that reached a transferable stage; Group C (N = 208): received the only 3 developing embryos. Clinical pregnancy, implantation, multiple pregnancy and delivery rates were registered. Kruskal-Wallis and Fisher Exact tests were applied as appropriate. RESULTS: Age and previous attempts were comparable in the 3 groups. Compared with Group A, Groups B and C had a higher oocyte recovery (10.7 ± 5.6 vs. 14.7 ± 8.0 vs. 13.8 ± 6.6), fertilization rate (75.97% vs. 81.60% vs. 83.29%) and percentage of embryos reaching a transferable stage on day 5 (39.98% vs. 63.99% vs. 60.97%), as well as a significantly higher clinical pregnancy (42.92% vs. 61.06% vs. 58.17%) and implantation rates (21.09% vs. 40.98% vs. 36.97%). The multiple pregnancy rate was higher in Groups B and C than in Group A (11.70% vs. 31.19% vs. 37.19%). The high order multiple pregnancy rate (> 2) was significantly increased in group C (1.06% vs. 0.92% vs. 14.05%). CONCLUSIONS: In patients with 3 or more day 5 developing embryos, delivery rates are similar if 2 or 3 embryos are transferred. The transfer of 3 embryos carries an unacceptable increase in the risk of high order multiple pregnancy, with its known consequences. According to our data, we should not exceed the number of 2 day-5 fresh embryos transferred.

Embryo Transfer Catheters: Softer is Easier.

OBJECTIVE: This study aimed to compare the use of semi-rigid and flexible catheters in terms of pregnancy rate and level of difficulty of the embryo transfer (ET) procedure. METHODS: Seven hundred and thirty-nine consecutive follicular aspirations for IVF or ICSI performed in our private fertility clinic within a 30-month period were randomly divided into two groups. Patients were allocated to either of the groups at the time of follicular retrieval with the aid of a random number generator according to the catheter used in ET: Group 1 (n = 370) - Frydman Classic Catheter 4.5 catheters; and Group 2 (n = 369) - Frydman Ultrasoft 4.5 catheters. Only patients offered ET in the randomized cycle were included (Group 1 = 363 and Group 2 = 340). Patients did not change groups after catheter allocation. Statistical analysis was carried out using an unpaired t-test or a chi square test as appropriate. Differences with P<0.05 were considered. RESULTS: A statistically significant difference in favor of the use of the Ultrasoft catheter was evident for parameters use of a tenaculum (19.8% vs. 5.9%) or a hysterometer (5.0% vs. 1.2%), presence of blood during post-ET inspection of the catheter (9.9% vs. 3.8%), and implantation rate (13.0% vs. 16.4%). Clinical pregnancy and delivery rates were comparable in both groups. CONCLUSION: Our results suggest that a softer catheter may help with difficult ETs. Softer catheters, as also reported by other authors, resulted in better implantation rates.

Vulvar carcinoma associated with lichen sclerosus. Experience at the Florence, Italy, Vulvar Clinic.

OBJECTIVE: To compare demographic and clinical characteristics of patients with lichen sclerosus (LS)-associated squamous cell carcinoma (SCC) of the vulva with those of patients with tumors not histologically associated with LS in a series of patients with vulvar SCC not HPV correlated. STUDY DESIGN: We retrospectively reviewed histologic specimens and clinical files of all vulvar SCCs referred to the Vulvar Clinic, University of Florence, Florence, Italy, since 1990. RESULTS: Twenty-five out of the 72 cases in this study (34.7%) were LS associated. Among these cases, 8 (32%) were diagnosed with LS before occurrence of the cancer and received treatment for the disease. In 17 cases the diagnosis of LS was simultaneous with that of SCC; in 13 cases the diagnosis was achieved by clinical examination and confirmed afterwards histologically. In 4 cases this was confirmed only by means of histologic examination. The shared profile of patients with LS-associated vulvar SCC was a subject (mean age, 72 years) seldom with a past medical history of vitiligo (16% of cases), with invasive cancer (92% of cases), clinically characterized by an exophitic tumor (73%), seldom ulcerated (18%) or showing hyperkeratosis (9%). Labia majora (32%), labia minora (27%) and vestibule (23%) were the most frequently involved sites. In most cases (80%) the cancer was limited to 1/3 of the vulvar region. An itch was the most frequent symptom. However, for all of these variables, no overall statistically significant difference was found with patients who had SCCs not associated with LS. CONCLUSION: The experience of the Vulvar Clinic, University of Florence, confirms the suggested role of LS as a possible precursor of vulvar carcinoma since 32% of our cases not HPV related were LS associated. We demonstrated that the profile of patients with LS-associated cancer does not differ from that of patients with cancer not associated with LS, excluding HPV-related cases. The existence of accessory conditions, probably needed to promote the progression from LS to cancer in a minority of subjects remains to be established.

Topical mometasone furoate for vulvar lichen sclerosus.

OBJECTIVE: To assess the efficacy and safety of treating vulvar lichen sclerosus with mometasone furoate, a synthetic analogue of beclomethasone. STUDY DESIGN: Thirty-one consecutive patients with biopsy-proven vulvar lichen sclerosus (LS) seen at the Florence Vulvar Clinic were entered into the study. Each subject's symptoms and the gross appearance of the lesions were recorded using a severity score according to the authors' previous studies. The patients were treated with a regimen of 0.1% mometasone furoate cream applications to the affected areas once daily for 4 weeks and then twice weekly for 8 weeks. After 12 weeks of treatment, the subjects were asked to rate the changes in their symptoms and were reevaluated by the same examiner for the gross appearance of the lesions. In every patient the before/after treatment total severity scores were compared using Wilcoxon's signed-rank test for statistical significance. RESULTS: With mometasone furoate all the patients had a significant improvement (P < .001) in the gross aspects of the disease and a very dramatic decrease in symptoms (P < .001), with nearly all the subjects having complete symptomatic remission. Compliance was excellent, and no side effects were observed. CONCLUSION: The improved risk/benefit ratio of mometasone furoate may minimize the possible long-term side effects expected when using a very potent topical steroid. Therefore, momentasone furoate cream may be an alternative to clobetasol propionate for treatment of vulvar LS, with similar efficacy but higher levels of safety and tolerability.

Development of a non invasive NGF-based therapy for Alzheimer's disease.

Nerve growth factor (NGF) deficits are linked to Alzheimer's Disease (AD), due to the role of NGF on basal forebrain cholinergic neurons (BFCN). We have further established that a disequilibrium in NGF signaling and/or processing from its precursor proNGF is also directly and causally related to the aberrant activation of an amyloidogenic route to neurodegeneration. The therapeutic potential of using human NGF to provide a long-lasting cholinergic trophic support, thereby preventing or slowing cognitive decline in AD patients, has therefore a strong rationale. However, a simple and practical means of delivering NGF to the brain in a safe and long-term manner, limiting the undesired adverse effects of NGF in activating nociceptive responses, has represented a significant challenge. For this reason, pilot clinical studies have been performed so far with invasive approaches requiring neurosurgery. We obtained a proof of principle, in neurodegeneration animal models, of an alternative, non-invasive delivery of NGF through an intranasal route, which facilitates access of NGF to the central nervous system (CNS), while minimizing the biodistribution of NGF to compartments where it activates undesired effects, such as pain. The ideal NGF product for a non invasive NGF-based therapy would be a recombinant NGF that, while exhibiting an identical biological activity to that of human NGF, can be traced, against the endogenous NGF, in order to optimize the therapeutical dose range and meet the required therapeutic window. We describe an engineered mutein of hNGF, hNGF-61, that is selectively recognized, against endogenous NGF, by a specific antibody. hNGF-61 mutein has an identical potency and bioactivity profile as hNGF, in vitro and in vivo. Moreover, hNGF-61 and hNGF are equally effective in rescuing the behavioral and neurodegenerative phenotype in adult and aged AD11 anti-NGF mice. Finally, we demonstrated that intranasally delivered hNGF-61 is significantly more effective than ocularly applied hNGF-61, to determine phenotypic rescue in AD11 mice. The development of hNGF-61 towards clinical applications in AD patients is under way.

Dermoscopic features of mucosal melanosis.

BACKGROUND: Melanosis (lentiginosis, labial melanotic macula) is a benign pigmented lesion of mucosa characterized by pigmentation of basal keratinocytes with melanocytic normal or slightly increased in number. Melanosis, particularly when occurring on genitalia, can clinically mimic mucosal melanoma thus creating concern in both the patient and the physician. OBJECTIVE: In this study dermoscopic features from a series of clinically equivocal (n=11) or clinically typical (n=10) mucosal melanosis were analyzed. METHODS: All the women consecutively seen at the Vulva Clinic of the Department of Obstetrics and Gynecology, University of Florence, Italy, from May 1, 2002 to June 30, 2002, were examined. RESULTS: Three major dermoscopic patterns were identified: (1) a "structureless" pattern, predominantly found in clinically equivocal vulvar melanosis, with a blue hue, associated with the presence of melanophages in the upper dermis, present in the majority of these lesions; (2) a "parallel pattern," often found in clinically typical melanotyc macules of the lips and penis; and (3) a "reticular-like" pattern associated with clinically equivocal melanosis occurring at peculiar sites such as the areola (all the three cases occurred at that site) or, rarely, on the lip. CONCLUSIONS: Dermoscopy can play a role in the noninvasive classification of mucosal melanosis. The risk of misclassification with melanoma is probably dependent on dermoscopy pattern shown by the lesion. Prospective studies including early melanomas are needed to establish diagnostic performance of dermoscopy in pigmented lesions of the mucosa.