Exosomal miRNA Biomarker Panel for Pancreatic Ductal Adenocarcinoma Detection in Patient Plasma: A Pilot Study.

Pancreatic ductal adenocarcinoma (PDAC) is rapidly becoming one of the leading causes of cancer-related deaths in the United States, and with its high mortality rate, there is a pressing need to develop sensitive and robust methods for detection. Exosomal biomarker panels provide a promising avenue for PDAC screening since exosomes are highly stable and easily harvested from body fluids. PDAC-associated miRNAs packaged within these exosomes could be used as diagnostic markers.

An Exosomal miRNA Biomarker for the Detection of Pancreatic Ductal Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) remains a difficult tumor to diagnose and treat. To date, PDAC lacks routine screening with no markers available for early detection. Exosomes are 40-150 nm-sized extracellular vesicles that contain DNA, RNA, and proteins.

Small extracellular vesicles in cancer.

Extracellular vesicles (EV) are lipid-bilayer enclosed vesicles in submicron size that are released from cells. A variety of molecules, including proteins, DNA fragments, RNAs, lipids, and metabolites can be selectively encapsulated into EVs and delivered to nearby and distant recipient cells. In tumors, through such intercellular communication, EVs can regulate initiation, growth, metastasis and invasion of tumors.

Exosomal biomarkers for cancer diagnosis and patient monitoring.

: In recent years, extensive research has been conducted on using exosomes as biomarkers for cancer detection. Exosomes are 40-150 nm-sized extracellular vesicles released by all cell types, including tumor cells. Exosomes are stable in body fluids due to their lipid bilayer member and often contain DNA, RNA, and proteins.

Mining Exosomal Genes for Pancreatic Cancer Targets.

Background: Exosomes, cell-derived vesicles encompassing lipids, DNA, proteins coding genes and noncoding RNAs (ncRNAs) are present in diverse body fluids. They offer novel biomarker and drug therapy potential for diverse diseases, including cancer.

Materials And Methods: Using gene ontology, exosomal genes database and GeneCards metadata analysis tools, a database of cancer-associated protein coding genes and ncRNAs (n=2,777) was established.

A new objective method for scoring human sperm hyperactivation based on head axis angle deviation.

The aims of the present study were, first to develop a new method for evaluating sperm hyperactivation (HA) based on sperm head axis angle deviation, and second to apply this method in scoring selected sperm populations during capacitation. This was made possible by improving our original superimposed image analysis system (SIAS). The new option on the system enables us to determine the real angular deviation of the entire head in successive superimposed frames.

Evaluation of the SQA IIB: a new version of a sperm quality analyzer.

Objective: To evaluate the SQA IIB (Sperm Quality Analyzer; MES-Medical Electronic Systems, Migdal Haemek, Israel), sperm analyzer that claims to read the three main sperm indices through algorithmic means and without a microscope.

Design: Controlled laboratory study comparing two methods of sperm analysis.

Setting: Academic environment.

Dynamic aspects concerned with the mechanism of separating motile sperm from nonmotile sperm, leukocytes, and debris with the use of high-density Percoll gradients.

Objective: To investigate the dynamics and the efficacy of separating motile from nonmotile sperm with the use of Percoll gradients as a function of centrifugation time.

Design: In vitro laboratory experiments on human sperm.

Setting: Male Infertility Institute.

Clinical, biochemical, and genetic findings in a large pedigree of male and female patients with 5 alpha-reductase 2 deficiency.

The present report describes a cluster of eight patients with male pseudohermaphroditism from a large pedigree with steroid 5 alpha-reductase 2 deficiency (5 alpha RD), who reside in Southern Lebanon. They were born with unambiguous female external genitalia and reared as girls until puberty, when masculinization occurred, followed by a change of gender role. Semen analysis and testicular histology revealed maturation arrest of spermatogenesis, with low sperm count and motility.

Evaluating the accuracy of different sperm counting chambers by performing strict counts of photographed beads.

Purpose: To suggest a method for evaluating the accuracy of different kinds of sperm counting chambers by eliminating errors concerned with human skills or semen properties. In this method, various concentrations were prepared from stocks of commercially available latex beads (Accu-beads, Hamilton-Thorn Research, Beverly, MA). Samples from identical preparations were loaded into different types of chambers, namely, hemocytometer (Neubauer, improved double, Superior Ltd, Germany), Makler (Sefi Medical Instruments, Haifa, Israel) and Horwell (Horwell Ltd, London, UK).

Inability of human sperm to change their orientation in response to external chemical stimuli.

Objective: To investigate whether human sperm can respond to external chemical stimuli by orienting themselves toward chemoattractants or withdrawing from hostile environments.

Design: Controlled laboratory assays.

Setting: Normal human sperm and two other flagellated micro-organisms were exposed to various potential chemoattractant or chemorepellent substances.

Sequential acquisition of chemotactic responsiveness by human spermatozoa.

Recent studies have indicated that human spermatozoa respond to follicular fluid by attraction to chemotactic factor(s) in the fluid, accompanied by enhancement of motility and ultimately hyperactivation. In this study, we quantified the sperm response. We exposed spermatozoa to a gradient of a chemotactically active fraction of follicular fluid (denoted as "the attractant") and separated the spermatozoa that accumulated in the attractant and those that did not.

Chemotaxis and chemokinesis of human spermatozoa to follicular factors.

Human spermatozoa accumulate in vitro in diluted follicular fluids obtained from follicles from which the eggs have been fertilized. Using capillary assays under a variety of experimental conditions (ascending or descending gradients of follicular fluid, or no gradient at all) and microscopic assays in which individual spermatozoa could be followed, we found that the sperm accumulation in follicular fluid was the result of both sperm chemotaxis and chemokinesis and eventually hyperactivation-like motility. We determined the optimal conditions for sperm accumulation, which involved sperm preincubation (possibly to induce sperm capacitation) and proper dilution of follicular fluid.

Atrial natriuretic peptide attracts human spermatozoa in vitro.

Here we report that atrial natriuretic peptide (ANP), a known activator of particulate guanylate cyclase, induces attraction and swimming speed enhancement of human spermatozoa in vitro. Using capillary assays under a variety of experimental conditions (ascending or descending gradients of ANP, or no gradient at all) and microscopic assays in which individual spermatozoa could be followed, we found that spermatozoa followed the gradient of ANP and accumulated in it. Speed enhancement was detected in the presence of ANP without a gradient.

Investigation in real time of the effect of gravitation on human spermatozoa and their tendency to swim-up and swim-down.

To investigate in real time if and how natural gravity affects rates of swim-up and swim-down of human spermatozoa, samples of motile or immobilized spermatozoa in a sealed mini-chamber were placed vertically on a 90 degrees tilted microscope. The mode of their sedimentation, as well as the difference in the rate of their swimming up and down, were observed directly over 30 min and analysed from photomicrographs. Under the influence of natural gravity force, most immobilized spermatozoa turned their heads down in about 5 min and then sank slowly at an average speed of 0.

Premature ovarian failure--the prognostic application of autoimmunity on conception after ovulation induction.

Objective: To assess whether the presence of autoimmune activity in patients with premature ovarian failure (POF) can predict the response to ovulation induction and conception.

Design: Assessment of autoimmune activity in patients with POF, correlating the response to ovulation induction with this autoreactivity.

Setting: Tertiary care academic center.

Use of a sealed minichamber for direct observation and evaluation of the in vitro effect of cigarette smoke on sperm motility.

Objective: To determine the in vitro effect of cigarette smoke on sperm motility and survival in an attempt to find a possible association with clinical studies that had pointed to the existence of such an effect in vivo.

Design: Laboratory experiments in which the effect of cigarette smoke on human sperm placed in a sealed minichamber was directly observed and determined by motion analysis.

Patients: High-quality fresh semen specimens were obtained from healthy donors and used for the experimental studies.

Use of a sealed mini-chamber to investigate human sperm motility in real time under aerobic and anaerobic conditions.

To study the correlation between metabolism and motility, ejaculated human spermatozoa were washed in media containing glucose, pyruvate, and deoxyglucose in various combinations. Spermatozoa suspended in these media were incubated in sealed mini-chambers and subjected to aerobic or anaerobic conditions at 37 degrees C. The effect on the patterns of sperm motility was investigated in real time by direct observation and objective determination with the multiple exposure photography (MEP) method.

A new model for investigating in real-time the existence of chemotaxis in human spermatozoa.

Objective: To suggest a new approach to the research of chemotaxis between various media and human spermatozoa to solve the riddle of its existence.

Design: Laboratory experiments in which chemotaxis between human spermatozoa and follicular fluid (FF) as well as N-formyl was investigated.

Setting: Male physiology laboratory and in vitro fertilization program unit.

Endogenous digoxin-like immunoreactivity measured in seminal fluid from a normal male population.

Endogenous digoxin-like immunoreactivity (EDLI) has been detected in different biological fluids and in several pathophysiological conditions. In this study, using radioimmunoassay we reported for the first time the existence of bound and unbound EDLI in normal seminal fluid. The unusual finding was the detection of unbound EDLI in the seminal fluid, while this reactivity was undetected in plasma.