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Luminal membrane retrieved after exocytosis reaches most golgi cisternae in secretory cells.

Dextran was used to trace membrane retrieved from the luminal surface after induced exocytosis in secretory cells of rat lacrimal and parotid glands. Two different approaches were used: (a) isolated acini were incubated in vitro with dextran followed by stimulation with carbamylcholine (lacrimal) or isoproterenol (parotid) and (b) rats were injected with isoproterenol followed by dextran infusion into the parotid duct in vivo. The main findings were the same regardless of the gland source or experimental approach. Dextran was taken up initially via coated pits into smooth-surfaced apical vesicles. Shortly thereafter it was found in multiple cell compartments: within the stacked Golgi cisternae, in condensing vacuoles, and in lysosomes. Uptake was more rapid and uniform in vivo; dextran was seen in multiple cisternae of numerous Golgi complexes within 5 min after infusion. In acini incubated in vitro uptake into Golgi cisternae was more delayed and occurred with increasing frequency up to 60 min; also, more dextran was taken up into lysosomes, which were more numerous in vitro than in vivo. The results demonstrate that, after exocytosis, membrane is removed from the cell surface via vesicles that fuse with multiple cell compartments. The two novel findings are: (a) the demonstration that the tracer can reach most of the Golgi cisternae in a given stack and (b) the demonstration of the rapidity with which the process takes place (i.e., within 5 min). The findings imply that at least some membrane retrieved from the cell surface after exocytosis fuses with the stacked Golgi cisternae.

Animals

WILDkCAT: extract, retrieve, and predict enzyme turnover numbers of constraint-based metabolic models.

SUMMARY: Accurate enzyme turnover numbers are essential for building enzyme-constrained genome-scale metabolic models. However, collecting and curating these parameters remains a major bottleneck. Indeed, kcat values are scattered across multiple databases, reported under varying experimental conditions, and often missing for many enzymes. To address this challenge, we present WILDkCAT, a Python-based pipeline that enables the retrieval of kcat values from wild-type enzyme measured under user-specified pH and temperature ranges for a given metabolic model. The application to Escherichia coli (iML1515) and Homo sapiens (Human-GEM) models demonstrated the ability of WILDkCAT to retrieve substantial kcat coverage and its applicability across diverse genome-scale models. AVAILABILITY AND IMPLEMENTATION: WILDkCAT is available at https://github.com/sysbiolux/WILDkCAT and from PyPI. WILDkCAT works on all major operating systems and computer architectures. The documentation is available at https://sysbiolux.github.io/WILDkCAT.

Software

Speed of retrieval from long-term memory in relation to age, familiarity, and datedness of information.

The time required to name four groups of pictures was measured in 83 community dwelling males (18-70 years of age). One group consisted of objects used 50 to 70 years ago (unique dated exemplars), and another consisted of objects unique to contemporary times (unique contemporary exemplars). For comparison, other pictures of contemporary and dated objects commonly used in both periods were employed. There were three major findings. One, the familiarity of the pictured object is the major determinant of the time required to retrieve its name. Older subjects named the dated unique objects that were relatively more familiar to them more rapidly than did younger adults, while the reverse was true for contemporary unique objects. No age difference in the speed of naming common contemporary objects was found. Two, the overall age-related difference in naming latency was attributed to perceptual-motor aspects of the task. Three, the physical features as well as the familiarity of the object contributed to the speed of retrieval of the object's name.

Adolescent

Acoustic signals in mouse maternal behavior: retrieving and cannibalism.

The purpose of this study was to show that juvenile acoustic signals are not essential prerequisites for inhibiting parental aggression or for eliciting maternal care. This was demonstrated by comparing the behavior of Rb-3 and GFF+/+ female mice with normal audition and that of a deaf mutant GFF dn/dn. 243 females and 609 litters were used to measure cannibalism and 54 females to measure speed of retrieval. Neither rate of cannibalism nor speed of retrieval were modified by deafness, thus verifying the proposed hypothesis.

Aggression

Oncologic research and computerized data retrieval.

A computer-based data retrieval system for clinical research in oncology is described. The system has been designed to enable busy clinicians to retrieve and analyze data directly without major involvement of a computer specialist. The system uses texts similar to those in daily practice. In most situations no code is printed. The other important feature of the system is its ability to analyze clinical data in the form of tables, frequency distribution, etc. It may also furnish actuarial characteristics of the total population or subpopulations. Mathematical functions are calculated easily. Amendments and updating are simple enabling follow-up data to be incorporated. The system is under use for about 2 years and appears to be a versatile tool in the field of clinical research.

Clinical Trials as Topic

Pre-Meta: priors-augmented retrieval for LLM-based metadata generation.

MOTIVATION: While high-throughput sequencing technologies have dramatically accelerated genomic data generation, the manual processes required for dataset annotation and metadata creation impede the efficient discovery and publication of these resources across disparate public repositories. Large language models (LLMs) have the potential to streamline dataset profiling and discovery. However, their current limitations in generalizing across specialized knowledge domains, particularly in fields such as biomedical genomics, prevent them from fully realizing this potential. This article presents Pre-Meta, an LLM-agnostic and domain-independent data annotation pipeline with an enriched retrieval procedure that leverages related priors-such as pre-generated metadata tags and ontologies-as auxiliary information to improve the accuracy of automated metadata generation. RESULTS: Validated using five selected metadata fields sampled across 1500 papers, the Pre-Meta assisted annotation experiment-without finetuning and prompt optimization-demonstrates a systemic improvement in the annotation task: shown through a 23%, 72%, and 75% accuracy gain from conventional RAG adoptions of GPT-4o mini, Llama 8B, and Mistral 7B respectively. AVAILABILITY AND IMPLEMENTATION: The code, data access, and scripts are available at: https://github.com/SINTEF-SE/LLMDap.

Metadata

Trophoblast Enrichment by Maternal Immune-Cell Depletion Using CD45 and CD56 Surface Markers in Trophoblast Retrieval and Isolation from the Cervix (TRIC).

Background: Trophoblast retrieval and isolation from the cervix (TRIC) has emerged as a promising alternative to invasive prenatal diagnostic procedures. However, contamination by maternal immune cells remains a major challenge that may compromise trophoblast purity and the reliability of downstream fetal genetic analyses. Methods: Maternal immune cells were selectively depleted by immunomagnetic sorting using antibodies targeting CD45 or CD56. The remaining cells were subsequently enriched for HLA-G-positive trophoblasts and characterized by immunofluorescence and gene-expression analyses using CD45, CD56, HLA-G, cytokeratin 7 (CK7), and β-human chorionic gonadotropin (β-hCG). Results: Compared with CD45-mediated depletion, CD56-mediated depletion demonstrated more efficient removal of maternal immune cells, as indicated by significantly reduced CD56 expression. CK7 expression showed an increasing trend following CD56 depletion, whereas β-hCG expression remained largely unchanged. Immunofluorescence analysis further demonstrated a significant increase in the proportion of CK7+/β-hCG+ trophoblast cells after CD56 depletion. Conclusions: Among the evaluated depletion strategies, CD56-mediated depletion demonstrated a more favorable profile for trophoblast-associated characteristics than CD45-mediated depletion, suggesting its potential contribution to further methodological optimization of trophoblast isolation in TRIC-based noninvasive prenatal genetic testing.

maternal immune cell

Rhinoplasty record and retrieval system.

The rhinoplastic surgeon must vary technique to fit the anatomic variations of nose and face. Our system to record and correlate preoperative findings with surgical methods could help accomplish this. It includes three forms to record pertinent findings of the history and physical examination, operative findings and techniques, and postoperative evaluations. These forms are numerically coded, and data can be transferred to a punch card system or computer cards for quick and easy retrieval and statistical analysis. The system tries to simplify the many anatomic and surgical variations, yet remain complete. We hope that this system, which has been used in other disciplines, can be applied to rhinoplasty. In this way, more scientific statistical data can be used to support surgical approaches.

Adult

Retrieving information on clinical methodology.

Evidence derived from several simple searchers of the literature suggests that one interested in identifying papers which discuss the methodologies of clinical trials will have reasonable success. However, the retrieval of a meaningful methodologic duscussion of any actual trial is unlikely because very little of this information has been published. Some suggestions to remedy this deficiency are given.

Clinical Trials as Topic

Effect of foot-shock intensity on amount of memory retrieval in rats by emotionally important stimuli in a drug-dependent learning escape design.

Drug-dependent learning (lack of transfer between drug states) was demonstrated and disrupted in an escape learning, forced choice T-maze task. A drug-dependent learning (DDL) group was trained to escape foot shock (0.65 mA) while in a drug (chlordiazepoxide hydrochloride) state. These rats subsequently responded randomly on non-shock test trials in the non-drug state, but continued to respond significantly (P less than 0.02) above random level when in the training drug state. Four transfer groups were also trained in the Drug state, but with a 1 kHz auditory tone simultaneously paired with foot shock. Each Transfer group received a different (0.10, 0.65, 3.5, and 4.5 mA) foot shock intensity during training. The auditory tone continued to be sounded during testing with no foot shock, and percentage correct turns, first-trial correct turns, and latency scores were significantly (P less than 0.01) different from the DDL group's performance. The results were interpreted as demonstrating that an emotionally-important auditory stimulus could initiate a memory retrieval process that could overcome a physiological state. This memory retrival process was not modified by wide variations in foot shock intensity.

Acoustic Stimulation

Disruption of drug-dependent learning (memory retrieval) using an ethanol drug state: a replication.

Eight groups (n = 8 per group) of rats were trained to escape foot shock by turning in the correct direction in a T-maze and 24 h after reaching criterion all groups were tested with no foot shock. The four experimental groups were given alternating drug and nondrug test days while the four control groups always experienced the same drug state in training and in testing. Two experimental groups (transfer) heard a 1 kHz tone that was simultaneously paired with foot shock during training, and the tone continued to be sounded on all test trials. These two groups responded significantly (P less than 0.05) above random level with low response latencies on all days of testing, regardless of drug state. The other two experimental groups (DDL) did not receive the tone, and these groups demonstrated dissociated performance. The present study, using 2400 mg/kg ethanol, replicated the DDL and memory retrieval results of our previous chlordiazepoxide studies.

Animals

Retrieval failures in alcohol state-dependent learning.

Retrieval processes subject to disruption by alteration of drug conditions were studied in a state-dependent learning experiment. On the first day of the 2-day experiment, 28 human subjects performed four verbal learning tasks in either an alcohol state (1.0 ml/kg) or a sober state, and the next day they attempted to recall the material in either the same or in an altered condition. Two of the tasks involved recall cues or prompts, and the other two required the subjects to recall the material unaided. Results indicated state-dependency on the two tasks that did not involve recall cues and no state-dependency on the two tasks involving cues. This finding indicates that memory failures resulting from a changed drug state can be reversed by appropriate experimental cueing or prompting, and implies that a learner's drug state has stimulus properties for recall. The role of drugs in contributing to a person's cognitive context and the resultant effects on memory are discussed.

Adolescent

The contingent negative variation during a memory retrieval task.

Evoked potentials were recorded from the human scalp during performance of a memory retrieval task modeled after a paradigm originated by Sternberg (1966). Subjects were required to decide whether a probe digit was contained in a series of one to four target digits presented a few seconds before. The amplitude of the contingent negative variation (CNV) preceding the probe digit and the speed of CNV resolution after the probe varied as a function of target set size. CNV amplitude was greatest when the set size was one. The smaller the set size, the more positive the evoked potential 300 msec after the probe, regardless of whether a motor response was required.

Adult

Ethanol and marihuana effects on event-related potentials in a memory retrieval paradigm.

Twelve men performed the Sternberg memory retrieval task under three experimental conditions: after oral doses of marihuana extract calibrated to contain 0.7 mg/kg delta9-tetrahydrocannabinol (THC), 1.0 ml/kg 95% ethanol, or placebo. Simultaneously, the EEG was recorded from Ca to linked ears and the EOG from leads above and below the right eye. In this task, subjects saw a set of 1 to 4 digits follwed by a warning tone that was followed 1.5 sec later by a test digit. Subjects indicated by pressing one of two buttons whether the test digit was in-set or out-of-the-set. There were no drug effects on N1 in the evoked potential to the warning tone, but P3 amplitude was smaller under THC and ethanol than under placebo. CNV amplitude in the interval between the warning tone and the test digit showed no drug effects, indicating that the subject was equally prepared for the test digit regardless of drug received. However, the latency of 50% resolution of the CNV was longer under THC than under placebo. THC also increased the reaction time for each set size by about 75 msec above the values for ethanol and placebo, the latter two not differing significantly. Set size affected N1 and P3 amplitudes and latencies and CNV amplitude, as well as 50% CNV resolution latency and reaction time, but there were no drug chi set size interactions.

Adolescent

Event-related potentials recorded from young and old adults during a memory retrieval task.

Six healthy old and 8 healthy young subjects each received a series of trials in a memory retrieval task devised by Sternberg (1966). On each trials, the subject received a memory set of 1-4 digits and was then shown a test digit. The subject's task was to press one of two response buttons indicating whether the test digit was a member of the memory set for that trial. Response time (RT) was found to be an increasing, linear function of the number of items held in memory. The slope of the RT function was a composite measure of the time necessary to process each additional item in memory while the intercept was a measure of stimulus encoding and response processes that do not depend on memory set size (see Sternberg 1966, 1969, 1975). We found that the latency of P3 to the test stimulus also increased with increases in memory set size, although the slope of the P3 was less than that for RT. We have suggested that the intercept of the P3 slope reflects the time it takes to encode the test stimulus before the evaluation of the stimulus starts, while the slope reflects the amount of time per digit needed to evaluate the set. We have suggested that the difference between RT and P3 slopes represents the additional time per digit which the subjects waits before making a response, due to low confidence occurring with more difficult task conditions (i.e., when set size = 4). We further suggest that the intercept of the RT-P3 latency slope is a reflection of pure response processes. Time estimates of these processes are made for young and old subjects.

Adult

A data storage and retrieval system for clinical research.

A data storage and retrieval system is presented that can be used for clinical research studies. The system is designed in such a way that the programs are independent of the application. It can be used by non-computer experts via a simple query language. Since it is bulit up in a modular way extensions can be easily made. The system is implemented on a PDP 11/70 under FORTRAN.

Computers

A computer-based obstetric data retrieval system.

A computer-based obstetric patient data retrieval system is described which permits physicians with no prior computer knowledge or experience to access a patient data base. The advantages are minimal physician instruction, opportunity to examine maternal and neonatal outcome of defined diagnostic or therapeutic subsets of the data base, and rapid recall of individual patients' data, without need for continuous assistance from computer specialists. Disadvantages are the costs and our inability to interface our medical information with hospital business office data. Two brief examples of use of the system are provided.

Computers

Long-term restoration of cardiac dystrophin expression in golden retriever muscular dystrophy following rAAV6-mediated exon skipping.

Although restoration of dystrophin expression via exon skipping in both cardiac and skeletal muscle has been successfully demonstrated in the mdx mouse, restoration of cardiac dystrophin expression in large animal models of Duchenne muscular dystrophy (DMD) has proven to be a challenge. In large animals, investigators have focused on using intravenous injection of antisense oligonucleotides (AO) to mediate exon skipping. In this study, we sought to optimize restoration of cardiac dystrophin expression in the golden retriever muscular dystrophy (GRMD) model using percutaneous transendocardial delivery of recombinant AAV6 (rAAV6) to deliver a modified U7 small nuclear RNA (snRNA) carrying antisense sequence to target the exon splicing enhancers of exons 6 and 8 and correct the disrupted reading frame. We demonstrate restoration of cardiac dystrophin expression at 13 months confirmed by reverse transcription-PCR (RT-PCR) and immunoblot as well as membrane localization by immunohistochemistry. This was accompanied by improved cardiac function as assessed by cardiac magnetic resonance imaging (MRI). Percutaneous transendocardial delivery of rAAV6 expressing a modified U7 exon skipping construct is a safe, effective method for restoration of dystrophin expression and improvement of cardiac function in the GRMD canine and may be easily translatable to human DMD patients.

Alternative Splicing