Coherence controlled holographic microscopy - a tool for detection of new biomarkers of head and neck squamous cell carcinoma.

Background: Squamous cell carcinoma of the head and neck is characterized by local invasiveness and metastases to regional lymph nodes. In 60% of cases, these tumours are dia-gnosed at an advanced stage, and the prognosis is unfavorable. One of the important factors of local, hematogenous or lymphogenic spread of the tumour in the human body is tumour cells migration ability.

Autopsy-Determined Atherosclerosis in Elite US Military Special Operations Forces.

Background: Autopsy studies of trauma fatalities have provided evidence for the pervasiveness of atherosclerosis in young and middle-aged adults. The objective of this study was to determine the prevalence of atherosclerosis in elite US military forces.

Methods: We conducted a retrospective study of all US Special Operations Command (USSOCOM) fatalities from 2001 to 2020 who died from battle injuries.

TGF β 3 immunoassay standardization: comparison of NIBSC reference preparation code 98/608 with avotermin lot 205-0505-005.

Juvista™ drug product contains human recombinant active transforming growth factor beta 3 (TGFβ3; avotermin). Juvista is being developed for the prevention and reduction of human scarring. Phase II and III clinical and development batches of Juvista were assayed for content by an immunoenzymometric assay (IEMA) using a National Institute for Biological Standards and Control (NIBSC) TGFβ3 reference material (98/608) and avotermin standard (Lot 205-0505-005).

Seasonal and inter-annual variation of Beryllium-7 deposition in birch-tree leaves and grass in the northeast upland area of the Czech Republic.

The activity concentrations of Beryllium-7 ((7)Be), a naturally occurring radioisotope produced in the atmosphere, were measured in leaves of birch-trees, above-ground parts of grass, soil and rainwater in the mountain massive Kralicky Sneznik (the northeast of the Czech Republic, altitude about 750 m) in the years of 2005, 2006 and 2007. Dried and ground samples of the plants and soils, and water samples from wet deposition were used to determine the (7)Be content using a semiconductor gamma spectrometer. The (7)Be values ranged from 147.

Production of fuculose-1-phosphate aldolase using operator-repressor titration for plasmid maintenance in high cell density Escherichia coli fermentations.

We report a novel application for the operator-repressor titration (ORT) plasmid maintenance system. The ability of ORT to maintain a plasmid during production of DNA has been demonstrated previously. In this study, we have used the ORT system to maintain a plasmid during high cell density cultivation and expression of a recombinant protein.

Antibiotic-free plasmid stabilization by operator-repressor titration for vaccine delivery by using live Salmonella enterica Serovar typhimurium.

Live, attenuated bacteria are effective vectors for heterologous antigen delivery. However, loss of heterologous gene-bearing plasmids is problematic, and antibiotics and their resistance genes are not desirable for in vivo DNA vaccine delivery due to biosafety and regulatory concerns. To solve this problem, we engineered the first vaccine delivery strain that has no requirement for antibiotics or other selectable marker genes to maintain the recombinant plasmid.

Effect of plasmid copy number and lac operator sequence on antibiotic-free plasmid selection by operator-repressor titration in Escherichia coli.

The Escherichia coli strain DH1lacdapD enables plasmid selection and maintenance that is free from antibiotics and selectable marker genes. This is achieved by using only the lac operator sequence as a selectable element. This strain is currently used to generate high copy number plasmids with no antibiotic resistance genes for use as DNA vaccines and for expression of recombinant proteins.

A modified Escherichia coli protein production strain expressing staphylococcal nuclease, capable of auto-hydrolysing host nucleic acid.

The large-scale production of recombinant biotherapeutics, particularly recombinant proteins, provides significant process and regulatory challenges to the biotechnology industry in order to meet the regulatory agencies stringent requirements in a cost-effective manner. Host cell derived nucleic acid causes problems from both a process and a regulatory perspective, as high molecular weight chromosomal DNA is responsible both for the viscosity of cell lysates, and it is a source of heterologous DNA sequences whose inclusion in the final product must be prevented. We have constructed a modified Escherichia coli JM107 expression host (JMN), containing a staphylococcal nuclease expression cassette, integrated into the host chromosome at the dif locus.

Pharmacy-based intervention to reduce antibiotic use for acute bronchitis.

Background: Intervention programs can reduce inappropriate antibiotic use for the treatment of acute bronchitis in a closed health maintenance organization model.

Objective: To evaluate the impact of a pharmacy-based intervention program intended to reduce antibiotic use in the treatment of acute bronchitis in a community-based physician group model.

Subjects: Adult and pediatric patients with an office or urgent care visit for acute bronchitis during the baseline and study periods were included in the study.

The impact of fluid-dynamic-generated stresses on chDNA and pDNA stability during alkaline cell lysis for gene therapy products.

Extensive tests have been carried out to assess the impact of fluid-dynamic-generated stress during alkaline lysis of Escherichia coli cells (host strain DH1 containing the plasmid pTX 0161) to produce a plasmid DNA (pDNA) solution for gene therapy. Both a concentric cylinder rheometer and two stirred reactors have been used, and both the alkaline addition and neutralization stages of lysis have been studied. Using a range of shear rates in the rheometer, stirrer speeds in the reactors, and different periods of exposure, their impact on chromosomal DNA (chDNA) and pDNA was assessed using agarose gel electrophoresis, a Qiagen Maxiprep with a polymerase chain reaction (PCR) assay, and a Qiagen Miniprep purification with a UV spectrophotometer.

Escherichia coli strains that allow antibiotic-free plasmid selection and maintenance by repressor titration.

We report the construction of two novel Escherichia coli strains (DH1lacdapD and DH1lacP2dapD) that facilitate the antibiotic-free selection and stable maintenance of recombinant plasmids in complex media. They contain the essential chromosomal gene, dapD, under the control of the lac operator/promoter. Unless supplemented with IPTG (which induces expression of dapD) or DAP, these cells lyse.

Purification of essentially RNA free plasmid DNA using a modified Escherichia coli host strain expressing ribonuclease A.

Regulatory agencies have stringent requirements for the large-scale production of biotherapeutics. One of the difficulties associated with the manufacture of plasmid DNA for gene therapy is the removal of the host cell-related impurity RNA following cell lysis. We have constructed a modified Escherichia coli JM107 plasmid host (JMRNaseA), containing a bovine pancreatic ribonuclease (RNaseA) expression cassette, integrated into the host chromosome at the dif locus.

Production of plasmid DNA for human gene therapy using modified alkaline cell lysis and expanded bed anion exchange chromatography.

We describe a process for the commercial manufacture of therapeutic grade plasmid DNA. The industrially scaleable unit operations employed in this process are: (i) optimized alkaline lysis; (ii) bag filtration; (iii) expanded bed anion exchange chromatography; (iv) ultrafiltration, and (v) size exclusion chromatography. These steps are scaleable alternatives to current approaches to plasmid DNA isolation such as high speed centrifugation for feed-stock clarification and solvent precipitation for plasmid concentration, and an efficient alternative to conventional low through-put packed bed chromatography.

Repressor titration: a novel system for selection and stable maintenance of recombinant plasmids.

The propagation of recombinant plasmids in bacterial hosts, particularly in Escherichia coli, is essential for the amplification and manipulation of cloned DNA and the production of recombinant proteins. The isolation of bacterial transformants and subsequent stable plasmid maintenance have traditionally been accomplished using plasmid-borne selectable marker genes. Here we describe a novel system that employs plasmid-mediated repressor titration to activate a chromosomal selectable marker, removing the requirement for a plasmid-borne marker gene.

[Trichorrhexis nodosa in electron microscopy].

Trichorrhexis nodosa, i. e. a hair fracturing into brush-like widened stumps, was analyzed in an imbecile 4 year-old boy without any sign of aminoaciduria or disturbance of copper metabolism.

[ECG similarities in the parents and offspring of thoroughbred horses].

The ECG characters were studied in two sires (Manrico and Infernal) and their 26-membered set of progeny as well as in one mare (Victoire) and her five daughters. The confer of some ECG characters from the sire's side as well as from the mare's side to the offspring was demonstrated. The consistency of some ECG characters was particularly obvious in externally dominant Manrico sire and his offspring as well as in the breeding mare and her five daughters (inclination of the electric cardiac axis, intrinsicoid deflexion lag, P wave shape, deep S in the 3rd connection).

[The effect of age and training on the ECG time curve of purebred horses].

In 246 horses of the English Thoroughbred breed of several age categories, divided into four test populations, the influence was studied of age and training on the pulse rate and time intervals of ECG (PQ, QRS, QT and QTc). It was found that with the increasing age and training action slowed down gradually. Duration of the atrioventricular path (PQ) is prolonged, time of ventricular activation (QRS) and time of the electrical systole (QT) including its value corrected according to the actual pulse rate (QTC).

[Single-lead bipolar electrocardiography in the diagnosis of heart diseases].

Sixty seven thoroughbred race horse of various age categories were examined electrocardiographically by bipolar leak-apex of the heart -- withers; length of training corresponded to the age categories. Basic time and voltage criteria of EKG were statistically evaluated according to electrocardiographic curves: TF = 35.1 = %/- 6.

[Distribution of surface electric potentials in the horse heart].

Surface electric potentials of the heart of four horses were measured by use of unipolar leak with the so called central Wilson's clip; the measurements were performed on 95 precisely fixed places of the equine body. Potentials found out in this way were re-measured with bipolar leaks: the negative electrode was fixed in the place of the highest negative potentials, the positive electrode was gradually attached to places with the highest values of positive potentials. The largest potential differences when the negative electrode was placed in the region of the heart were obtained from the region of the withers, spinal and scapular regions, from forehead and nose.