Molecular cloning of MER-2, a human chromosome-11-encoded red blood cell antigen,using linkage of cotransfected markers

We report the molecular cloning of a human gene MER-2located on chromosome 11 that encodes a cell surface antigen which is polymorphic on red blood cells. An essential element of the cloning strategy was cotransfection-induced linkage of pSV2-neo, which encodes resistance to the antibiotic G418, to the human MER-2gene. An important feature of the pSV2-neo construct is that the same gene (the transposon, Tn5) that encodes G418 resistance in eukaryotic cells confers neomycin resistance in bacteria. Chinese hamster ovary (CHO) cells were cotransfected with pSV2-neo and genomic DNA from a CHO ×human cell hybrid containing a single human chromosome (chromosome 11). Transfectants expressing both the human MER-2gene and G418 resistance were isolated by selection in the antibiotic G418, followed by indirect immunofluorescence using the monoclonal antibody 1D12, which recognizes the MER-2 antigen, manual enrichment, and single-cell cloning. Genomic DNA from a primary transfectant positive for MER-2expression and G418 resistance was used to construct a cosmid library and cosmid clones able to grow in neomycin were isolated. Of 150,000 cosmid clones screened, 90 were resistant to neomycin and of these, 11 contained human repetitive sequences. Five neomycin-resistant cosmid clones containing human repetitive DNA were able to transfect CHO cells for G418 resistance and MER-2expression.     

Utility of a multiplex PCR assay for detecting herpesvirus DNA in clinical samples

A multiplex PCR was designed to amplify herpes simplex virus types 1 and 2, cytomegalovirus, and varicella-zoster virus DNA present in a diverse range of clinical material. The susceptibility of these viruses to in vivo inhibition by at least one antiviral drug was an important consideration in their inclusion in the multiplex detection system. An aliquot of equine herpesvirus was introduced into each specimen prior to extraction and served as an indicator of potential inhibitors of the PCR and a detector of suboptimal PCR conditions. Compared to virus isolation and immunofluorescence-based antigen detection, the multiplex assay yielded higher detection rates for all viruses represented in the assay. The turnaround time for performance of the assay was markedly reduced compared to those for the other techniques used to identify these viruses. More than 21,000 tests have been performed using the assay. Overall, the multiplex PCR enabled the detection of substantially increased numbers of herpesviruses, in some cases in specimens or anatomical sites where previously they were rarely if ever identified using traditional detection methods.     

High rate of transfer of Staphylococcus aureus from parental skin to infant gut flora

Many Swedish infants carry Staphylococcus aureus in their intestinal microflora. The source of this colonization was investigated in 50 families. Infantile S. aureus strains were isolated from rectal swabs and stool samples at 3 days and at 1, 2, 4, and 8 weeks of age. The strains were identified by using the random amplified polymorphic DNA method and compared to strains from swab cultures of the mothers' hands, nipples, and nares and from the fathers' hands and nares. Maternal stool samples were also obtained at a later stage to compare infant and adult intestinal S. aureus colonization. Although 60% of 1-month-old children had S. aureus in the stools, this was true of only 24% of the mothers. The median population numbers in colonized individuals also differed: 10(6.8) CFU/g of feces among infants at 2 weeks of age versus 10(3.2) CFU/g of feces in the mothers. Of S. aureus strains in the stools of 3-day-old infants, 90% were identical to a parental skin strain. A total of 96% of infants whose parents were S. aureus skin carriers had S. aureus in their feces and 91% had the same strain as at least one of the parents. In comparison, only 37% of infants to S. aureus-negative parents had S. aureus in the stool samples. Thus, infantile intestinal S. aureus colonization was strongly associated with parental skin S. aureus carriage (P = 0.0001). These results suggest that S. aureus on parental skin establish readily in the infantile gut, perhaps due to poor competition from other gut bacteria.     

HLA-A2-restricted CD8+-cytotoxic-T-cell responses to novel epitopes in Mycobacterium tuberculosis superoxide dismutase, alanine dehydrogenase, and glutamine synthetase

Major histocompatibility complex class I-restricted CD8(+) cytotoxic T lymphocytes (CTL) are implicated in protective Th1 immunity to Mycobacterium tuberculosis infection. We report the identification of three novel HLA-A*0201-restricted CTL epitopes within mycobacterial superoxide dismutase (SodA), L-alanine dehydrogenase (AlaDH), and L-glutamine synthetase (GlnS) proteins.     

Specific binding proteins for human chorionic gonadotrophin in a patient with trophoblastic disease

Human chorionic gonadotrophin (hCG) is present in high concentration in patients with trophoblastic disease. The hCG concentration parallels tumor activity. The first known case of an endogenous binder for hCG in the serum of a patient with trophoblastic disease is reported. The patient's hCG concentration was not detectable when analyzed by a radioimmunoassay using a double antibody method; however, it was positive by a radioimmunoassay using polyethylene glycol precipitation. A high serum blank was also observed in this patient. This led the authors to suspect the presence of an endogenous binder for hCG. Scatchard analysis of the patient's serum revealed a high affinity binder with Ka = 1 x 10(10) M(-1), concentration of binding site = 2 x 10(-11) M. This investigation strongly suggested a specific binder(s) for hCG in the serum of this patient.     

In vitro performance of the novel coronary sinus AutoRetroPerfusion Cannula

Myocardial salvage through coronary sinus intervention has been documented. The AutoRetroPerfusion Cannula is a novel device that is able to perfuse the coronary bed retrogradely through the coronary sinus with arterial blood generated from a peripheral artery with no need for a pump. The cannula consists of a distal end that, once secured in the coronary sinus, opens an umbrella-like membrane to create pressure in the coronary sinus, and at the same time has small channels directed backwards to the right atrium to provide pressure relief. The cannula is introduced from the axillary vein under local anesthesia and the proximal end, which consists of a graft, is anastomosed to the axillary artery to start autoperfusion once the distal end is secured in the coronary sinus and the occluding membrane is open. The AutoRetroPerfusion Cannula was tested in the in vitro mock loop under 50-120 mm Hg of proximal pressure and 50, 100, and 150 ml/min of total flow in the cannula. We were able to achieve the nominal design point of 40-80 mm Hg of distal pressure and 50-150 ml/min of distal flow by adjusting the number, diameter, and length of the small backwards channels.     

Use of ECT to Treat Bipolar Depression in a Mental Retardate with Cerebral Palsy

We report the use of electroconvulsive therapy (ECT) for treatment of a bipolar episode in a mildly retarded young man with cerebral palsy. Difficulties in the diagnosis and treatment of affective disorders in the retarded are discussed.     

Subclasses of event-related potentials: response-locked and stimulus-locked components

The manner in which information processing proceeds and the relationship of the components of the cerebral "event-related" evoked potential (ERP) to the presumed stages of such processing are uncertain. We studied these relationships by recording long-latency evoked potentials from the scalp in conjunction with electromyographic (EMG) recording in experiments in which subjects were required to make a motor response to one of two auditory stimuli. The cerebral response was averaged from the tone onset (stimulus-synchronized average) and the EMG onset (response-synchronized average). We found that N2 was quite distinct from the other ERP components (P165 and P3) in its lack of a close relationship to the motor response required in a sensory discrimination task. This suggests that, contrary to general belief, the P165-N2-P3 components of the ERP are not generated in an orderly sequence, but rather are generated separately and are possibly related to different cognitive processes.     

Patient Satisfaction with AbobotulinumtoxinA for Aesthetic Use in the Upper Face: A Systematic Literature Review and Post-hoc Analysis of the APPEAL Study

BACKGROUND: AbobotulinumtoxinA (AboBoNT-A; Dysport^®; Ipsen, Boulogne-Billancourt, France/Azzalure^®; Galderma, Lausanne, Switzerland) is a botulinum neurotoxin type A approved for aesthetic use in the treatment of glabellar lines in adult patients under 65 years in Europe, the United States, and other countries. OBJECTIVE: We sought to analyze current literature on patient satisfaction with aboBoNT-A for upper facial aesthetic indications. METHODS: A systematic review of literature databases (PubMed/MEDLINE, Embase, the Cochrane Library, and Google Scholar) was performed to identify English-language publications reporting on patients with aesthetic indications (including glabellar lines and wrinkles) receiving aboBoNT-A, that assessed patient and/or physician satisfaction with treatment, with no restrictions on comparator studies. Structured data extraction was used to enable inter-study analysis. A post-hoc analysis was also performed to assess patient satisfaction by sex and age, using results from the noninterventional APPEAL study of patients’ satisfaction with aboBoNT-A for treating glabellar lines. RESULTS: Overall, 22 original research papers were identified. Patient satisfaction rates for aboBoNT-A treatment were significantly higher versus placebo from two weeks to between three and five months postinjection. At two to three weeks postinjection, patient satisfaction rates were 52% and 99% across studies. In studies with later time points, patient satisfaction rates were 85 to 87 percent at 5 months and between 25 and 100 percent at 6 months post-injection. Physician satisfaction was also high (97%–100%, across three treatments). No notable differences in patient satisfaction by sex or age were observed in the APPEAL study. CONCLUSION: High rates of patient satisfaction have been achieved with aboBoNT-A treatment for upper facial aesthetic indications. Despite the current recommended interval of ≥12 weeks, satisfaction with the aesthetic results of aboBoNT-A therapy is still evident up to 6 months post-injection in some patients.