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Background: Patients with infantile fibrosarcoma (IFS) have shown strong and long-lasting responses to larotrectinib, a tropomyosin receptor kinase inhibitor (TRKi), in single-arm clinical trials. Conventional chemotherapy has also shown important efficacy. But, until now, no comparative data exist. This study aims to assess the therapeutic benefit of larotrectinib over the current standard of care (SOC) of chemotherapy in paediatric patients with locally advanced or metastatic IFS., Patients and Methods: EPI VITRAKVI is a retrospective, observational, externally controlled study (NCT05236257). Data of patients aged ≤21 years with locally advanced or metastatic IFS treated with larotrectinib in the phase I/II SCOUT trial (NCT02637687) were compared with those of an external historical control group (data of Institut Curie and Cooperative Weichteilsarkom Studiengruppe) treated with a chemotherapy-based regimen. Between-group differences were assessed after balancing groups using inverse probability of treatment weighting (IPTW)., Results: In total, 93 patients were compared, 51 in the larotrectinib arm and 42 in the external control arm. After therapy, 4 patients (7.8%) in the larotrectinib group had a medical treatment failure event [start of new systemic treatment (2 cases), mutilating surgery (2 cases)] versus 15 (35.7%) in the external control group [start of new systemic treatment (6 cases), mutilating surgery (5 cases), radiation therapy (2 cases), and death (2 cases)]. Larotrectinib was associated with an 80% reduced likelihood of encountering a medical treatment failure event, when compared to the external control group (weighted and stratified hazard ratio 0.20, 95% confidence interval 0.06-0.63, P = 0.0060). These results were confirmed by sensitivity analyses, including exact matching, and subgroup analyses for number of lines of treatment., Conclusions: Treatment with larotrectinib reduced the need of subsequent therapies compared to SOC with chemotherapy in children with locally advanced or metastatic IFS, regardless of the line of treatment., Competing Interests: Disclosure SKK, MF, and MK are Bayer employees. DO had a consultant activity for Bayer and Roche and is an Independent Data Monitoring Committee (IDMC) member for a Lilly product and is a consultant for Novartis Pharma France and Eusapharm. MC had a consultant activity for Bayer. MSS had advisory activity for Bayer, Roche, and Sobi. All other authors have declared no conflicts of interest., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Introduction: Cephalomedullary nails (CMN) have proven to be effective for the fixation of unstable pertrochanteric hip fractures, but limited data exists regarding the degree of shortening that occurs with nailing systems that offer "dynamically" versus "statistically" locked helical blades. This study aimed to determine the incidence, timing, and amount of postoperative fracture shortening in patients with pertrochanteric hip fractures treated with "dynamically" versus "statistically" locked helical blades., Methods: A retrospective review was performed on patients requiring operative fixation of pertrochanteric fractures. Inclusion criteria were patients aged 55 or older, low energy mechanism of injury, primary surgical open reduction and internal fixation of the injury, surgery performed by a fellowship-trained orthopaedic traumatologist, and radiographic follow up until healing or failure (minimum of 3 months). Quantification of fracture shortening was determined using the lateral screw protuberance method. Fracture shortening measurements were calculated from immediate post-operative films and subsequent radiographs at each follow-up visit., Results: 222 patients met inclusion criteria (123 using Depuy Synthes Trochanteric Fixation Nail proximal femoral nailing system (TFN) with static locking; 65 TFN with dynamic locking; 23 using Depuy Synthes Trochanteric Fixation Nail-Advanced proximal nailing system (TFNA) with static locking; and 11 TFNA with dynamic locking). When comparing the final screw protuberance, the TFN dynamic (10.31 mm), TFNA static (9.52 mm), and TFNA dynamic (7.02 mm) groups were found to be significantly less compared to the TFN static cohort (11.67 mm; p < 0.001). The distribution of fracture shortening was found to be significant (p < 0.01). Interval measurements of fracture shortening by repeated measures of ANOVA within groups showed statistically significant increases in screw protuberance for the TFN static (p < 0.001), TFN dynamic (p < 0.001), and TFNA static (p < 0.01) cohorts within the first 60 days postoperatively. The TFNA with dynamic locking group was found to have statistically significant increases until the first 100 days post-operatively (p < 0.02)., Discussion: The incidence of fracture shortening remains high even with the use of cephalomedullary nails, regardless of dynamization status. Current pertrochanteric fixation constructs continue to favor fracture healing at the direct expense of fracture shortening., Competing Interests: Declaration of Competing Interest The authors report no conflicts related to this manuscript., (Copyright © 2022. Published by Elsevier Ltd.)

Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.

Research Question: What are the most influential articles in reproductive biology journals from 1980 to 2019 according to Altmetric Attention Score (AAS), number of citations and Relative Citation Ratio (RCR)?, Design: Cross-sectional study of reproductive biology articles indexed in the National Institutes of Health Open Citation Collection from 1980 to 2019. Data were downloaded on 20 May 2021. The 100 articles with highest AAS, RCR and number of citations were analysed., Results: Twenty-one reproductive biology journals were identified, including 120,069 articles published from 1980 to 2019. In total 227 reproductive biology classics were identified due to some overlap between the three lists. Compared with the 100 articles with the highest AAS (after excluding articles featured on both lists), the 100 top-cited articles were older (2014 versus 2001, mean difference [95% confidence interval] 13.5 [11.5, 15.5]), less likely to be open access (64% versus 85%), more likely to be reviews (42% versus 12%) and less likely to be observational studies (9% versus 51%) and randomized clinical trials (0% versus 5%). These same trends were observed in analyses comparing the 100 articles with highest AAS to the 100 articles with highest RCR. The most common topic was assisted reproduction, but prominent topics included infertility for top AAS articles, reproductive technology in animals for top-cited articles, and polycystic ovary syndrome for top RCR articles., Conclusions: Formerly, influential articles in reproductive biology journals were evaluated by absolute citation rates and subject to limitations of conventional bibliometric analysis. This is the first comprehensive study to use altmetrics and citation-based metrics to identify reproductive biology classics., (Copyright © 2022. Published by Elsevier Ltd.)

Acute myeloid leukemia (AML) is an attractive entity for the development of chimeric antigen receptor (CAR) T-cell immunotherapy because AML blasts are susceptible to T-cell-mediated elimination. Here, we introduce sialic acid-binding immunoglobulin-like lectin 6 (Siglec-6) as a novel target for CAR T cells in AML. We designed a Siglec-6-specific CAR with a targeting domain derived from the human monoclonal antibody JML-1. We found that Siglec-6 is commonly expressed on AML cell lines and primary AML blasts, including the subpopulation of AML stem cells. Treatment with Siglec-6 CAR T cells confers specific antileukemia reactivity that correlates with Siglec-6 expression in preclinical models, including induction of complete remission in a xenograft AML model in immunodeficient mice (NSG/U937). In addition, we confirmed Siglec-6 expression on transformed B cells in chronic lymphocytic leukemia (CLL), and specific anti-CLL reactivity of Siglec-6 CAR T cells in vitro. Of particular interest, we found that Siglec-6 is not detectable on normal hematopoietic stem and progenitor cells (HSPCs) and that treatment with Siglec-6 CAR T cells does not affect their viability and lineage differentiation in colony-formation assays. These data suggest that Siglec-6 CAR T-cell therapy may be used to effectively treat AML without the need for subsequent allogeneic hematopoietic stem cell transplantation. In mature normal hematopoietic cells, we detected Siglec-6 in a proportion of memory (and naïve) B cells and basophilic granulocytes, suggesting the potential for limited on-target/off-tumor reactivity. The lack of expression of Siglec-6 on normal HSPCs is a key to differentiating it from other Siglec family members (eg, Siglec-3 [CD33]) and other CAR target antigens (eg, CD123) that are under investigation in AML, and it warrants the clinical investigation of Siglec-6 CAR T-cell therapy., (© 2021 by The American Society of Hematology.)

Objectives: While India has made substantial progress in introducing new vaccines and scaling up immunization coverage, inequities persist sub-nationally. This study was performed to investigate the risk of under-immunization based on class membership and to identify heterogeneous classes based on sociodemographic characteristics in pediatric and maternal populations in India through latent class analysis., Methods: Data from the most recent National Family Health Survey conducted in 2015-2016 were used. Latent class analysis was used to model immunization coverage in children aged 12-23 months and mothers, and to identify subgroups to characterize those at risk of not being immunized., Results: Patterns of sociodemographic characteristics were found to contribute to non-immunization or under-immunization among pediatric and maternal populations in India. Individuals who fit into one of three categories were identified in both populations: those at high, medium, and lower risk of not being immunized. Lower socioeconomic status, lack of antenatal care, and lower maternal education put individuals at higher risk of not being immunized with routine childhood vaccines and maternal tetanus toxoid., Conclusions: Predisposing risk factors can persistently impact immunization status despite improvements in immunization access in India. Tailored programmatic interventions should be developed to improve immunization coverage among those children and mothers who are at highest risk of being under-immunized or not immunized., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

During platelet spreading, the actin cytoskeleton undergoes rapid rearrangement, forming filopodia and lamellipodia. Controversial data have been published on the role of lamellipodia in thrombus formation and stability. The Wiskott-Aldrich syndrome protein-family verprolin-homologous protein (WAVE)-regulatory complex, which has been shown in other cells to drive lamellipodium formation by enhancing actin nucleation via the actin-related protein 2/3 (Arp2/3) complex, is activated by Ras-related C3 botulinum toxin substrate 1 (Rac1) interaction with the WAVE complex subunit cytoplasmic fragile X mental retardation 1-interacting protein 1 (Cyfip1). We analyzed Cyfip1flox/floxPf4-Cre mice to investigate the role of Cyfip1 in platelet function. These mice displayed normal platelet counts and a slight reduction in platelet volume. Activation of mutant platelets was only moderately reduced to all tested agonists as measured by αIIbβ3 integrin activation and P-selectin surface exposure. However, lamellipodium formation of mutant platelets was completely abolished on different matrices. Nevertheless, Cyfip1-deficient platelets formed stable thrombi on collagen fibers ex vivo and in 2 models of occlusive arterial thrombosis in vivo. Similarly, the hemostatic function and maintenance of vascular integrity during inflammation of the skin and lung were unaltered in the mutant mice. Investigation of platelet morphology in an induced thrombus under flow revealed that platelets rather form filopodia in the thrombus shell, and are flattened with filopodium-like structures when in direct contact to collagen fibers at the bottom of the thrombus. We provide for the first time direct evidence that platelet lamellipodium formation is not required for stable thrombus formation, and that morphological changes of platelets differ between a static spreading assay and thrombus formation under flow., (© 2019 by The American Society of Hematology.)

This study investigates potential limitations of 1,3-propanediol formation by Lactobacillus diolivorans. Particular focus is given to enhanced glycerol utilization as well as the elimination of by-product formation. The key aspect is a modulation of the redox household by process engineering through the application of carbon pulses. A shift in the product pattern of C3 products was achieved, improving the ratio of 1,3-propanediol versus 3-hydroxypropionic acid up to a level of 20:1. Moreover, in combination with alternative feeding strategies this ratio was enhanced up to 45:1 and the maximum observed productivity of 1,3-propanediol could be significantly increased to 1.7g/Lh., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Coronary artery fistula (CAF) is an incidental finding in up to 0.2% of cardiac catheterization studies and is a rare pathologic condition affecting up to 1% of the general population. We report two cases of CAF diagnosed by coronary angiography and managed by surgical ligation, with excellent postoperative outcome. Anatomic location, clinical presentation, investigation, and treatment of CAF are discussed., (Copyright © 2015 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Microorganisms appear as ideal catalysts for chemical conversions. Diverse metabolic routes seem to open doors to the whole range of chemistry. Indeed, a vast amount of scientific papers suggesting new microbial cell factories for old and new products is published every year. However, only very few of them reached industrial relevance. Chemical balances and some metabolic tricks allow natural microorganisms the efficient production of some chemicals, but not others. So first of all it is important to choose metabolically feasible products of value for synthetic chemistry. Here we see a clear task for the chemical and biotechnology industries to communicate for defining the right target molecules. Finally, despite our limited current knowledge, synthetic biology points to a future independent from natural strain backgrounds., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Integration of two fluorescence imaging methods enables tracking of the formation of fibrillar Aβ peptide amyloid aggregates in neurons, as discussed by Esbjörner and colleagues in this issue of Chemistry & Biology. This approach has the potential to fundamentally improve our understanding of the onset and therapeutic intervention of neurodegenerative diseases., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A subgroup of severe combined immunodeficiencies (SCID) is characterized by lack of T and B cells and is caused by defects in genes required for T- and B-cell receptor gene rearrangement. Several of these genes are also involved in nonhomologous end joining of DNA double-strand break repair, the largest subgroup consisting of patients with T(-)B(-)NK(+)SCID due to DCLRE1C/ARTEMIS defects. We postulated that in patients with ARTEMIS deficiency, early and late complications following hematopoietic cell transplantation might be more prominent compared with patients with T(-)B(-)NK(+)SCID caused by recombination activating gene 1/2 (RAG1/2) deficiencies. We analyzed 69 patients with ARTEMIS and 76 patients with RAG1/2 deficiencies who received transplants from either HLA-identical donors without conditioning or from HLA-nonidentical donors without or with conditioning. There was no difference in survival or in the incidence or severity of acute graft-versus-host disease regardless of exposure to alkylating agents. Secondary malignancies were not observed. Immune reconstitution was comparable in both groups, however, ARTEMIS-deficient patients had a significantly higher occurrence of infections in long-term follow-up. There is a highly significant association between poor growth in ARTEMIS deficiency and use of alkylating agents. Furthermore, abnormalities in dental development and endocrine late effects were associated with alkylation therapy in ARTEMIS deficiency.

Natural killer (NK) cells are a major component of the anti-tumor immune response. NK cell dysfunctions have been reported in various hematologic malignancies, including chronic lymphocytic leukemia (CLL). Here we investigated the role of tumor cell-released soluble and exosomal ligands for NK cell receptors that modulate NK cell activity. Soluble CLL plasma factors suppressed NK cell cytotoxicity and down-regulated the surface receptors CD16 and CD56 on NK cells of healthy donors. The inhibition of NK cell cytotoxicity was attributed to the soluble ligand BAG6/BAT3 that engages the activating receptor NKp30 expressed on NK cells. Soluble BAG6 was detectable in the plasma of CLL patients, with the highest levels at the advanced disease stages. In contrast, NK cells were activated when BAG6 was presented on the surface of exosomes. The latter form was induced in non-CLL cells by cellular stress via an nSmase2-dependent pathway. Such cells were eliminated by lymphocytes in a xenograft tumor model in vivo. Here, exosomal BAG6 was essential for tumor cell killing because BAG6-deficient cells evaded immune detection. Taken together, the findings show that the dysregulated balance of exosomal vs soluble BAG6 expression may cause immune evasion of CLL cells.

Super-resolution fluorescence imaging can provide insights into cellular structure and organization with a spatial resolution approaching virtually electron microscopy. Among all the different super-resolution methods single-molecule-based localization microscopy could play an exceptional role in the future because it can provide quantitative information, for example, the absolute number of biomolecules interacting in space and time. Here, small organic fluorophores are a decisive factor because they exhibit high fluorescence quantum yields and photostabilities, thus enabling their localization with nanometer precision. Besides past progress, problems with high-density and specific labeling, especially in living cells, and the lack of suited standards and long-term continuous imaging methods with minimal photodamage render the exploitation of the full potential of the method currently challenging., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Familial hemophagocytic lymphohistiocytosis (FHL) is a genetically determined hyperinflammatory syndrome caused by uncontrolled immune response mediated by T-lymphocytes, natural killer (NK) cells, and macrophages. STXBP2 mutations have recently been associated with FHL5. To better characterize the genetic and clinical spectrum of FHL5, we analyzed a cohort of 185 patients with suspected FHL for mutations in STXBP2. We detected biallelic mutations in 37 patients from 28 families of various ethnic origins. Missense mutations and mutations affecting 1 of the exon 15 splice sites were the predominant changes detectable in this cohort. Patients with exon 15 splice-site mutations (n = 13) developed clinical manifestations significantly later than patients with other mutations (median age, 4.1 year vs 2 months) and showed less severe impairment of degranulation and cytotoxic function of NK cells and CTLs. Patients with FHL5 showed several atypical features, including sensorineural hearing deficit, abnormal bleeding, and, most frequently, severe diarrhea that was only present in early-onset disease. In conclusion, we report the largest cohort of patients with FHL5 so far, describe an extended disease spectrum, and demonstrate for the first time a clear genotype-phenotype correlation.

Targeted irradiation of the bone marrow with radiolabeled monoclonal antibodies (radioimmunotherapy) represents a novel therapeutic approach with both myeloablative and antileukemic potential. In an open-label, single-center pilot study, 30 pediatric and adolescent patients undergoing hematopoietic cell transplantation for malignant (n = 16) and nonmalignant (n = 14) disorders received treatment with a ⁹⁰Y-labeled anti-CD66 monoclonal antibody. Patients with a high risk of relapse (n = 7) received additional treatment with standard conditioning based on either total body irradiation or busulfan to intensify the antileukemic effect. In patients with comorbidities (n = 23), radioimmunotherapy was combined with a reduced-intensity conditioning regimen to reduce systemic toxicity. Preferential irradiation of the bone marrow was achieved in all patients. Nonrelapse mortality was 4 (13%) of 30 patients. In patients with malignant diseases, the probabilities of overall and disease-free survival at 2 years were 0.69 (95% confidence interval 0.37-0.87) and 0.46 (95% confidence interval 0.19-0.70), respectively. In patients with nonmalignant diseases, the probability of both overall and disease-free survival at 2 years was 0.94 (95% confidence interval 0.63-0.99). This pilot study demonstrates that radioimmunotherapy is effective in achieving myeloablation with low additional toxicity when used in combination with standard or reduced-intensity conditioning in young patients.

Background: Little is known about the comparative effect of weight-loss diets on metabolic profiles during dieting., Objective: The purpose of this study was to compare the effect of a low-carbohydrate diet (< or =20 g/d) with a high-carbohydrate diet (55% of total energy intake) on fasting and hourly metabolic variables during active weight loss., Design: Healthy, obese adults (n = 32; 22 women, 10 men) were randomly assigned to receive either a carbohydrate-restricted diet [High Fat; mean +/- SD body mass index (BMI; in kg/m(2)): 35.8 +/- 2.9] or a calorie-restricted, low-fat diet (High Carb; BMI: 36.7 +/- 4.6) for 6 wk. A 24-h in-patient feeding study was performed at baseline and after 6 wk. Glucose, insulin, free fatty acids (FFAs), and triglycerides were measured hourly during meals, at regimented times. Remnant lipoprotein cholesterol was measured every 4 h., Results: Patients lost a similar amount of weight in both groups (P = 0.57). There was an absence of any diet treatment effect between groups on fasting triglycerides or on remnant lipoprotein cholesterol, which was the main outcome. Fasting insulin decreased (P = 0.03), and both fasting (P = 0.040) and 24-h FFAs (P < 0.0001) increased within the High Fat group. Twenty-four-hour insulin decreased (P < 0.05 for both groups). Fasting LDL cholesterol decreased in the High Carb group only (P = 0.003). In both groups, the differences in fasting and 24-h FFAs at 6 wk were significantly correlated with the change in LDL cholesterol (fasting FFA: r = 0.41, P = 0.02; 24-h FFA: r = 0.52, P = 0.002)., Conclusions: Weight loss was similar between diets, but only the high-fat diet increased LDL-cholesterol concentrations. This effect was related to the lack of suppression of both fasting and 24-h FFAs.

Herein, we present an unusual case of a 48-year-old patient with Ehlers-Danlos syndrome (classic type) with multiple cardiovascular pathologies, including mitral regurgitation, a dilated aortic root, and chronic atrial fibrillation. A complex mitral valve repair, modified cryo-Maze procedure, and aortic valve-sparing operation (reimplantation) were performed, which were further complicated by the patient's fragile tissues. The patient was discharged on postoperative day 10 in good health with normal mitral and aortic valve function. We believe that this is the first report of such an extensive cardiac procedure in a patient with Ehlers-Danlos syndrome., (2010 The Society of Thoracic Surgeons. Published by Elsevier Inc. All rights reserved.)

Increasing use of preimplantation genetic diagnosis (PGD) poses numerous clinical, social, psychological, ethical, legal and policy dilemmas, many of which have received little attention. Patients and providers are now considering and using PGD for a widening array of genetic disorders, and patients may increasingly seek 'designer babies.' In the USA, although governmental oversight policies have been discussed, few specific guidelines exist. Hence, increasingly, patients and providers will face challenging ethical and policy questions of when and for whom to use PGD, and how it should be financed. These issues should be better clarified and addressed through collection of data concerning the current use of PGD in the USA, including factors involved in decision making about PGD use, as well as the education of the various communities that are, and should be, involved in its implementation. Improved understanding of these issues will ultimately enhance the development and implementation of future clinical guidelines and policies.

Adenosine deaminase (ADA) deficiency is a systemic metabolic disease that causes an autosomal recessive variant of severe combined immunodeficiency (SCID) and less consistently other complications including neurologic abnormalities. Hematopoietic stem cell transplantation (HSCT) is able to correct the immunodeficiency, whereas control of nonimmunologic complications has not been extensively explored. We applied HSCT in 15 ADA-deficient patients consecutively treated at our institutions since 1982 and analyzed long-term outcome. Seven patients received transplants without conditioning from HLA-matched family donors (MFDs); the other 8 patients received conditioning and were given transplants either from HLA-mismatched family donors (MMFDs; n = 6) or from matched unrelated donors (MUDs; n = 2). At a mean follow-up period of 12 years (range, 4-22 years), 12 patients are alive with stable and complete immune reconstitution (7 of 7 after MFD, 4 of 6 after MMFD, and 1 of 2 after MUD transplantation). Six of 12 surviving patients show marked neurologic abnormalities, which include mental retardation, motor dysfunction, and sensorineural hearing deficit. We were unable to identify disease or transplantation-related factors correlating with this divergent neurologic outcome. The high rate of neurologic abnormalities observed in long-term surviving patients with ADA deficiency indicates that HSCT commonly fails to control CNS complications in this metabolic disease.

Proteins have evolved to fold and function within a cellular environment that is characterized by high macromolecular content. The earliest step of protein folding represents intrachain contact formation of amino acid residues within an unfolded polypeptide chain. It has been proposed that macromolecular crowding can have significant effects on rates and equilibria of biomolecular processes. However, the kinetic consequences on intrachain diffusion of polypeptides have not been tested experimentally, yet. Here, we demonstrate that selective fluorescence quenching of the oxazine fluorophore MR121 by the amino acid tryptophan (Trp) in combination with fast fluorescence correlation spectroscopy (FCS) can be used to monitor end-to-end contact formation rates of unfolded polypeptide chains. MR121 and Trp were incorporated at the terminal ends of polypeptides consisting of repetitive units of glycine (G) and serine (S) residues. End-to-end contact formation and dissociation result in "off" and "on" switching of MR121 fluorescence and underlying kinetics can be revealed in FCS experiments with nanosecond time resolution. We revisit previous experimental studies concerning the dependence of end-to-end contact formation rates on polypeptide chain length, showing that kinetics can be described by Gaussian chain theory. We further investigate effects of solvent viscosity and temperature on contact formation rates demonstrating that intrachain diffusion represents a purely diffusive, entropy-controlled process. Finally, we study the influence of macromolecular crowding on polypeptide chain dynamics. The data presented demonstrate that intrachain diffusion is fast in spite of hindered diffusion caused by repulsive interactions with macromolecules. Findings can be explained by effects of excluded volume reducing chain entropy and therefore accelerating the loop search process. Our results suggest that within a cellular environment the early formation of structural elements in unfolded proteins can still proceed quite efficiently in spite of hindered diffusion caused by high macromolecular content.

optomotor-blind (omb) and optomotor-blind related-1 (org-1) encode T-domain DNA binding proteins in Drosophila. Members of this family of transcription factors play widely varying roles during early development and organogenesis in both vertebrates and invertebrates. Functional specificity differs in spite of similar DNA binding preferences of all family members. Using a series of domain swap chimeras, in which different parts of OMB and ORG-1 were mutually exchanged, we investigated the relevance of individual domains in vitro and in vivo. In cell culture transfection assays, ORG-1 was a strong transcriptional activator, whereas OMB appeared neutral. The main transcriptional activation function was identified in the C-terminal part of ORG-1. Also in vivo, OMB and ORG-1 showed qualitative differences when the proteins were ectopically expressed during development. Gain-of-function expression of OMB is known to counteract eye formation and resulted in the loss of the arista, whereas ORG-1 had little effect on eye development but caused antenna-to-leg transformations and shortened legs in the corresponding gain-of-function situations. The functional properties of OMB/ORG-1 chimeras in several developmental contexts was dominated by the origin of the C-terminal region, suggesting that the transcriptional activation potential can be one major determinant of developmental specificity. In late eye development, we observed, however, a strong influence of the T-domain on ommatidial differentiation. The specificity of chimeric omb/org-1transgenes, thus, depended on the cellular context in which they were expressed. This suggests that both transcriptional activation/repression properties as well as intrinsic DNA binding specificity can contribute to the functional characteristics of T-domain factors.

Reproductive capacity in women declines dramatically beyond the fourth decade of life. Oocyte quality seems to be the primary determinant of reproductive potential, although age-related uterine changes may also contribute. Underlying reasons for reproductive decline in women remain unclear, and both ovarian and neuroendocrine mechanisms have been proposed.A number of age-related endocrinologic changes precede menopause and predict diminished reproductive capacity. Thus, "ovarian reserve" screening may identify patients in whom attempts at conventional assisted reproduction is warranted before proceeding with ovum donation. Techniques to preserve the maternal genetic contribution to offspring (in lieu of egg donation), including germinal vesicle transfer and donor ooplasm, are under investigation.

A human immunodeficiency virus-discordant couple failed to conceive through in vitro fertilization and intracytoplasmic sperm injection of cryopreserved semen banked by the human immunodeficiency virus-positive partner. The husband subsequently had acquired immunodeficiency syndrome and died. The subsequent transfer of cryopreserved embryos resulted in pregnancy. Both mother and child are human immunodeficiency virus negative.

Objective: Originating from the pituitary gland, TSH secretion is regulated predominantly by thyroid-releasing hormone (TRH) neurons located in the hypothalamus. Norepinephrine and dopamine have important effects in modulation of TSH secretion. An inhibitor of catecholamine synthesis, alpha-methyl-para-tyrosine (AMPT) has been used in several studies of the regulation of human TSH secretion. The short-term effects (<8 hours) of low doses of AMPT include stimulation of pituitary TSH secretion by selective lowering of brain dopamine levels. After prolonged administration of AMPT (>24 hours), theoretically both dopamine and norepinephrine levels are lowered significantly in the brain, although this has not been reported previously., Methods: Nine subjects (five women and four men) received a total of five 1-g doses of AMPT or five 50-mg doses of promethazine (active placebo) over 28 hours in a randomized, double-blind, placebo-controlled crossover design in which the active and control tests were separated by 4-6 weeks. Blood samples were obtained over 24 hours (18 time points) on day 2 of each condition., Results: Changes in prolactin secretion and 6-hydroxymelatonin sulfate excretion indirectly showed the effects of AMPT on dopamine and norepinephrine. The typical circadian rhythm of TSH secretion was blunted by AMPT throughout the night; at ten time points, the difference between the two groups was statistically significant (P <.01). The long-term effects of repeated doses of AMPT were inhibition of TSH secretion and significant attenuation of the circadian rhythm of TSH. Additionally, AMPT induced low norepinephrine levels, which counteracted the stimulatory effect of low dopamine levels on TSH., Conclusion: Through its inhibitory effect on TRH, norepinephrine appeared to be involved in the regulation of TSH.

A review of 1000 aspirations of oocyte donors was performed. Only witnessed events necessitating hospitalization or emergent intervention were considered significant. Seven (0.7% incidence) cases were noted, including severe ovarian hyperstimulation syndrome (n = 3), adverse reaction to intravenous anesthesia (n = 2), intra-abdominal bleeding after aspiration (n = 1), and bladder atony with hematuria after aspiration (n = 1).

The first capillary array scanner for time-resolved fluorescence detection in parallel capillary electrophoresis based on semiconductor technology is described. The system consists essentially of a confocal fluorescence microscope and a x,y-microscope scanning stage. Fluorescence of the labelled probe molecules was excited using a short-pulse diode laser emitting at 640 nm with a repetition rate of 50 MHz. Using a single filter system the fluorescence decays of different labels were detected by an avalanche photodiode in combination with a PC plug-in card for time-correlated single-photon counting (TCSPC). The time-resolved fluorescence signals were analyzed and identified by a maximum likelihood estimator (MLE). The x,y-microscope scanning stage allows for discontinuous, bidirectional scanning of up to 16 capillaries in an array, resulting in longer fluorescence collection times per capillary compared to scanners working in a continuous mode. Synchronization of the alignment and measurement process were developed to allow for data acquisition without overhead. Detection limits in the subzeptomol range for different dye molecules separated in parallel capillaries have been achieved. In addition, we report on parallel time-resolved detection and separation of more than 400 bases of single base extension DNA fragments in capillary array electrophoresis. Using only semiconductor technology the presented technique represents a low-cost alternative for high throughput DNA sequencing in parallel capillaries.

Imidocarb dipropionate, formulated as Imizol, is used for the treatment and prophylaxis of bovine babesiosis. Several studies have shown that imidocarb remains detectable in edible ovine and bovine tissues for several months after dosing but the mechanism of retention remains unknown. In this study, the mechanism of imidocarb retention was investigated by measuring the binding of [14C]imidocarb to bovine hepatocytes, erythrocytes, sub-cellular fractions and isolated bovine macromolecules. The proportion of [14C]imidocarb (10 microM) bound to cells in suspension culture (1 x 10(7) cells.ml-1) was found to be substantially greater to hepatocytes (56.5%) than to erythrocytes (4.6%). Studies with washed erythrocytes reconstituted in plasma indicated that approximately 70% of the [14C]imidocarb was bound to plasma proteins, 10% to erythrocytes, and 20% remained free. Measurement of [14C]imidocarb binding to sub-cellular fractions prepared from bovine liver revealed preferential accumulation in the nuclear, rather than in the mitochondrial, microsomal or cytosolic fractions. Binding capacities of selected bovine macromolecules for [14C]imidocarb were in the order deoxy-ribonucleic acid (DNA) = ribonucleic acid (RNA) > > alpha 1-acid glycoprotein (AGP) > serum albumin (BSA) > haemoglobin (Hb). DNA binding sites for imidocarb remained unsaturated over the concentration range 0-100 microM [14C]imidocarb. Competitive binding studies between imidocarb and pentamidine or spermidine provided evidence for common DNA binding sites. These studies indicated that preferential binding of [14C]imidocarb to hepatocytes compared with erythrocytes observed in vitro was a result of substantial reversible binding to nucleic acids and that the same cellular mechanism may be implicated in the slow elimination of imidocarb from edible tissues in vivo.

A triplet pregnancy occurred in a 51-year-old menopausal women after oocyte donation. Three of the four transferred embryos implanted, and the patient elected to carry the pregnancy. She delivered three viable infants weighing between 1500 and 2100 gm at 33 weeks' gestation by cesarean section.

Objective: Our purpose was to examine the effect of the recipient's age on pregnancy and miscarriage rates after oocyte donation., Study Design: A retrospective analysis of 307 consecutively performed donor oocyte cycles was undertaken. Recipients were divided into two groups: younger group < or = 42 years old (165 cycles) versus older group > 42 years old (145 cycles). Pregnancy outcomes between groups were compared. Confounding variables such as donor and cycle characteristics, recipient characteristics, and male parameters were also examined., Results: Both groups had the same incidence (6%) of failed fertilization cycles. At least one embryo was transferred in the remaining 286 cycles. Ongoing or delivered pregnancy rates per embryo transfer were similar for younger and older groups (30.2% vs 30.6%). Differences in miscarriage rates were not statistically significant (9.8% vs 16.3%). There were no differences in the cycle parameters related to donor age, number of oocytes aspirated, number of oocytes fertilized, and number of embryos transferred. Donor sperm was more frequently utilized in the older group (19 vs 4 cycles, p = 0.0002). In the remaining cycles male partners of older recipients were significantly older (37.9 +/- 0.5 years vs 43.9 +/- 0.6 years, p < 0.0001), but there were no differences in semen parameters or fertilization rates. Older recipients were more likely to be parous but also to have experienced a previous miscarriage., Conclusion: Recipient age does not adversely affect cycle outcome with donor oocytes. This implies that aging of the uterus is not of clinical significance to patients electing this method of infertility treatment.

The age at which the uterus no longer provides a receptive environment for embryo implantation and development remains unknown. We assessed whether menopausal women between the ages of 50 and 59 years could be prepared for pregnancy with oocyte donation. All potential recipients underwent extensive screening to ensure that they were in excellent physical, reproductive, and psychological health before enrollment. Of 18 patients initially screened (mean [SD] age 52.2 years [2.5]), 14 couples were entered into the study. Oocytes were donated by fertile women (28.1 [2.7] years) who provided gametes after pituitary downregulation with leuprolide acetate and routine ovarian hyperstimulation by human menopausal gonadotropin. 22 donor follicle aspirations resulted in 21 embryo transfers to 14 recipients. Pregnancies were established in 9 women. 1 pregnancy ended in a preclinical loss. The ratio of clinical pregnancies per transfer attempt was 38% (8/21), with an implantation rate per transferred embryo of 19%. Of the 8 clinical pregnancies, 1 spontaneous abortion occurred at 7 weeks' gestation. 3 women have delivered, 2 at 35 weeks' by emergency caesarean section for worsening preeclampsia and 1 at 37 weeks' after a normal vaginal birth. 4 pregnancies continue to progress normally beyond the second trimester. Women may become pregnant after the age of natural menopause by means of in-vitro fertilisation of donated oocytes. However, thorough medical screening before embryo transfer is essential to reduce to a minimum obstetric risks common in this older population.

A severe pelvic infection resulting in a tuboovarian abscess after transcervical embryo transfer is reported. The case is unique in that the recipient was an agonadal woman who had not undergone prior transvaginal aspiration. Although rare, pelvic infection after embryo transfer may occur in spite of normal precautions.

The gonadotropin-releasing hormone antagonist offers several advantages over the use of the agonist and allows several physiologic questions to be addressed. In this study, we evaluated the ability of Nal-Glu to acutely inhibit the luteinizing hormone surge and prevent ovulation. We also assessed whether recovery of the follicle would be possible after several days of gonadotropin deprivation and estradiol decrement. Eight normal ovulatory women were randomized to control or Nal-Glu-treated cycles (50 micrograms/kg intramuscularly) for 3 to 4 days. Monitoring was carried out with daily vaginal ultrasonographic scans and serum estradiol levels and twice-daily serum luteinizing and follicle-stimulating hormone levels. Nal-Glu acutely inhibited the luteinizing hormone surge and ovulation, even when administered as late as the onset of the luteinizing hormone surge. Evidence was provided that spontaneous follicular rescue recurred in eight of 10 cycles after 3 to 4 days of Nal-Glu administration. Although an estradiol to follicular size dissociation occurred with Nal-Glu, subsequent ovulation occurred in 5.1 +/- 0.6 days after the last Nal-Glu dose. The decrement in estradiol after Nal-Glu administration correlated negatively with the days required for subsequent ovulation to occur (r = 0.77, p less than 0.05). The subsequent luteal phase also was normal in terms of length and progesterone levels. These data confirm the potency and efficacy of Nal-Glu in acutely inhibiting gonadotropins and extends our knowledge on the physiologic characteristics of the dominant follicle.

The agonistic effect of the gonadotropin-releasing hormone agonist often necessitates an extended period of treatment, resulting in a longer treatment cycle and increased cost. We have evaluated the intermittent use of a gonadotropin-releasing hormone antagonist, Nal-Glu, and have designed a new, simplified protocol for its use in in vitro fertilization. Seven women who had previously undergone treatment with leuprolide acetate and human menopausal gonadotropins were treated with Nal-Glu. Leuprolide acetate, 1 mg/day subcutaneously, was administered in the midluteal phase until down regulation was achieved (estradiol less than 30 pg/ml). Human menopausal gonadotropins, three to four ampules per day intramuscularly, was administered in conjunction with 500 micrograms subcutaneous leuprolide acetate. In the treatment cycles Nal-Glu (50 micrograms/kg/day) was administered intramuscularly on cycle day 1 or 2 for 3 days to achieve down regulation. Human menopausal gonadotropins, three to four ampules intramuscularly, was then administered daily without the antagonist. Nal-Glu was resumed when the follicles reached 14 to 16 mm and was continued until the day of human chorionic gonadotropin administration. Compared with leuprolide acetate-human menopausal gonadotropins cycles, the days required for down-regulation with Nal-Glu were significantly shortened (20.6 +/- 4.1 vs 1.6 +/- 0.3 days, p less than 0.001), as was total cycle length (31.3 +/- 5.8 vs 11.0 +/- 1.0 days, p less than 0.01). The mean number of days of treatment with human menopausal gonadotropins, the mean number of ampules of human menopausal gonadotropins, peak estradiol levels, the number of oocytes, and the percent of oocytes fertilized were not statistically different. No luteinizing hormone surges were detected with Nal-Glu in serum or urine. Nal-Glu was well tolerated, and five pregnancies have resulted. We conclude that intermittent administration of Nal-Glu is highly effective in achieving down-regulation and blocking spontaneous luteinizing hormone surges. Compared with leuprolide acetate-human menopausal gonadotropins cycles, an equally high oocyte and embryo yield may be anticipated. This new protocol substantially decreases cycle length and increases patient convenience.

In the clinical practice of human in vitro fertilization, pregnancy is dependent on embryo implantation. Pregnancy is a function of the number of embryos transferred, with multiple embryos resulting in a higher likelihood of pregnancy. We formulated a mathematic model of embryo implantation. This model describes embryo implantation as dependent on three factors--transfer efficiency, embryo quality, and endometrial receptivity. Application of existing embryo implantation data to this model allows the calculation of the approximate value of each of these factors. On the basis of historic data, data obtained from our in vitro fertilization program, and these theoretic considerations, it is our hypothesis that (1) there is an inherent inefficiency associated with the mechanical transfer of embryos into the uterine cavity, which limits the maximal embryo implantation rate; (2) the quality of embryos produced by controlled ovarian hyperstimulation, follicle aspiration, and in vitro fertilization is very high and approaches that of embryos produced in natural cycles in vivo; and (3) endometrial receptivity is markedly diminished in stimulated cycles and is the current rate-limiting step of pregnancy success of in vitro fertilization.

The utilization of oocyte and preembryo donation in the treatment of infertility has recently expanded. Since 1983, modifications in the retrieval of donated female gametes have enhanced both the safety and the efficacy of the procedure. Pregnancy rates in women receiving donated preembryos are remarkably high, with success occurring in 25% to 50% of transfer cycles. The technology used for preembryo donation continues to evolve, and the current and future role ovum donation in the treatment of infertility.

Culdocentesis has been used routinely in the evaluation of ectopic pregnancy. To determine whether culdocentesis continues to play an important role, we reviewed the operative findings of 297 women undergoing an operative procedure because of ectopic pregnancy. Culdocentesis was performed before surgery in 252 cases. Of those, 210 (83%) had positive test results and 42 (17%) had negative results. Positive results accurately predicted a ruptured ectopic pregnancy in 50% of cases, whereas negative test results were predictive of an unruptured ectopic pregnancy in 58% of cases. Six patients without an ectopic pregnancy underwent unnecessary laparotomy because of positive culdocentesis test results, whereas 27 patients who were discharged from the emergency room after negative test results were obtained subsequently were found to have ectopic pregnancies. The results of this study imply that culdocentesis is of limited value in a clinical setting in which sensitive and rapid testing and pelvic ultrasonography are used.