Abnormalities in proliferation and protein synthesis in skin fibroblast cultures from patients with diabetes mellitus

Several aspects of in-vitro cell growth and protein synthesis were assessed in cultures of skin fibroblasts from subjects with juvenile-onset diabetes mellitus (JODM) or adult-onset diabetes mellitus (AODM) and from age-matched nondiabetic controls (C). There was an inverse correlation between increasing age and both the log-phase doubling rate and saturation density at confluence in C fibroblasts. JODM and AODM cells had a reduction in both indices of cell population growth in comparison with age-matched C fibroblasts. Fibroblasts grown in the presence of 0.3 micronM hydrocortisone were stimulated to grow more rapidly and to a greater saturation density. Stimulation of cell division by hydrocortisone accentuated the abnormalities in growth of JODM and AODM fibroblasts. Total protein and collagen synthesis was measured whtn the fibroblasts had grown to confluency in medium with or without hydrocorticone. Hydrocorticone did not produce a significant change in total protein and collagen synthesis per cell by C fibroblasts. Fibroblasts from AODM had a 180 per cent increase in total protein and collagen synthesis in the presence of hydrocortisone. In contrast, total protein and collagen synthesis decreased 40 per cent in fibroblasts from JODM when grown in the hydrocortisone medium. These studies indicate that skin fibroblast cultures from patients with diabetes exhibit abnormalities in cell proliferation. Furthermore, hydrocortisone appears to unmask diffeerences in protein synthesis that distinguish JODM and AODM fibroblasts in culture.     

Spectrophotometric determination of diphenhydramine. HC1 in pharmaceutical preparations

A new spectrophotometric method has been developed for determining diphenhydramine. HC1, based on solvent extraction into chloroform of the complex formed with bromocresol green. The complex solution in chloroform showed maximum absorption at 415 nm and obeyed Beer's law over the concentration range of 3.0-12.0 microgram/ml. The molar absorptivity of the complex was 2.02 x 10(4). Complex formation and extraction was complete and quantitative over the pH range from 2 to 5. The ratio of diphenhydramine to bromocresol green was 1:1. Excipients, coloring matter, flavoring agents, and other substances likely to be present in diphenhydramine preparations do not interfere in the determination. Direct determinations in tablet, capsule, sirup, and lotion preparations were carried out satisfactorily, and the average recovery was 100 +/- 1.0%.     

Reliability of phase-velocity measurements of tibial bone

BACKGROUND AND PURPOSE: The Bone Stiffness Measurement Device-Swing is capable of measuring the propagation velocity of flexural waves in human tibial bone, which relates to bending stiffness. If the interrater and intrarater reliability of measurements obtained with the device are established, it can be used with confidence in assessing changes in bone. The purposes of this study were to detect potential sources of measurement error and to establish the interrater and intrarater reliability of measurements taken with the device. SUBJECTS AND METHODS: In the first part of the study, a random-effects design was used to obtain phase-velocity measurements in subjects without known orthopedic or neurological impairments. The second part of the study consisted of possible applications of the device with mixed designs on subjects with spinal cord injuries. By means of generalizability theory, multiple sources of error (eg, occasion, clinician, repetition) were estimated. For the clinical trial, 17 persons with spinal cord injuries not older than 5 weeks were tested. RESULTS: The standard error of measurement (SEM) for intrarater reliability measurements ranged from 7.3 to 9.8 m x s(-1) . The SEMs for interrater reliability measurements ranged from 5.7 to 9.5 m x s(-1). The SEMs for measurements obtained by a single clinician in a clinical population ranged from 11.9 to 39.7 m x s(-1). CONCLUSION AND DISCUSSION: The reproducibility of measurements obtained with the device is suitably high for the device to be used for evaluation in clinical and research settings.     

Comparison between enzyme-linked immunosorbent assay and cytotoxic cross-match procedures for detecting IgG anti-donor antibodies

BACKGROUND: Disadvantages inherent to complement-dependent cytotoxicity cross-match (CDC XM) methods are the requirements for complement and viable target cells, detection of antibodies (Abs) against non-HLA antigens, and subjective scoring. Cross-Stat (SangStat Medical Corp., Menlo Park, CA), a recently developed enzyme-linked immunosorbent assay XM procedure for the detection of IgG anti-donor HLA Abs, is theoretically devoid of these flaws. METHODS: We compared results of Cross-Stat and our standard anti-human globulin (AHG)-enhanced CDC XM procedure on 524 sera from 230 transplant candidates, which were evaluated against 51 cadaveric donors. RESULTS: There was a significant correlation between AHG-CDC IgG XM and Cross-Stat results (P<0.001). For false negative sera, repeat AHG-CDC IgG XMs were still positive after platelet absorption, indicating that the Abs present were either non-HLA Abs or anti-HLA class II. Flow cytometry testing of false positive sera usually (42/62) substantiated Cross-Stat results, indicating that the discrepancy with AHG-CDC IgG XM is caused by greater sensitivity of Cross-Stat. Relative to the AHG-CDC XM, the sensitivity of Cross-Stat was 100%, the specificity was 93%, the positive predictive value was 73%, and the negative predictive value was 100%. A technical shortcoming of the Cross-Stat assay is that the frequency of indeterminate samples in the assays was 15%. Among 49 Cross-Stat negative vs. 13 Cross-Stat positive primary cadaveric renal allograft recipients (all AHG-CDC IgG-XM negative), there was no statistical difference in overall graft survival. CONCLUSION: Given the important theoretical advantages of enzyme-linked immunosorbent assay-based XM methods over the CDC XM, however, further testing of the clinical relevance of the Cross-Stat is warranted.     

Pattern formation in chick feather development: distribution of beta 1-integrin in normal and scaleless embryos

We have examined the immunolocalization of beta 1-integrin during feather development in the spino-lumbar tract of the backskin from normal and scaleless chick embryos. beta 1-integrin appears during early feather development in three distinct phases which correspond to important developmental events. The first phase (5-5 1/2 days of incubation; Hamburger and Hamilton [H.H.] stage 27) represents the period prior to the formation of dermis. During this phase, beta 1-integrin antiserum labels mesenchymal cells located in the central region of the spino-lumbar tract where the initiation site for feather development is located. The second phase (5 1/2-7 1/2 days of incubation; H.H. stages 28-32) corresponds to the period during which dermis is formed. The cells that make up the dermis are readily distinguished by their lack of beta 1-integrin immunostaining. The third phase (7 1/2-10 days of incubation; H.H. stages 33-36) begins with the sudden appearance of beta 1-integrin in the central and lateral regions of the dermis. The pattern of beta 1-integrin immunostaining in scaleless backskin becomes different from that of normal backskin during this phase. In normal backskin the dermal condensations of feather germs are not labeled with the beta 1-integrin antiserum. This produces a heterogeneous immunostaining pattern very similar to the pattern seen for Type I collagen (Mauger et al. [1982] Dev. Biol. 94:93-105). In contrast, homogeneous immunostaining is observed in the dermis of scaleless backskin. The initial time of appearance, manner of appearance, and pattern of integrin expression in the third phase suggest that beta 1-integrin may be involved in the stabilization of the feather pattern. We also observed the appearance of beta 1-integrin on the epidermal basal cells during the time of feather follicle formation. The beta 1-integrin antiserum reacts strongly with the baso-lateral surfaces of normal basal cells, yet the basal surfaces of the scaleless basal cells are unstained. This lack of immunostaining along the basal surfaces of the scaleless basal cells may relate to the abnormal adhesion between the epidermis and dermis in scaleless backskin.     

Dietary modulation of human platelet phenolsulphotransferase activity

1. The mammalian phenolsulphotransferase enzymes are known to play a major role in both the detoxification and possibly the activation of pre-carcinogenic phenols and aromatic amines. 2. Vegetable cytosol preparations were tested in vitro for their ability to affect the sulphation of two reference compounds (rho-nitrophenol and dopamine, which are selective substrates for the phenol and monoamine forms of phenolsulphotransferase respectively), and to act as substrates for the enzymes in comparison with the same reference compounds. 3. The majority of cytosols greatly decreased (> 80%) the sulphation of either or both the reference compounds. This effect may have been due to either enzyme inhibition or substrate binding. 4. Whereas some of the cytosols were sulphated under the assay conditions, most were not. Additionally, it was found that a cytosol that decreased the sulphation of the two reference compounds was not necessarily poorly sulphated itself. 5. It is concluded that dietary factors have the potential to play a major role in modulating the sulphation detoxification pathway, and have wide ranging implications with regard to adverse drug reactions.     

The logic of Ménage à Trois

Recent studies of socially monogamous species have shown that in many cases females do not copulate exclusively with their pair mates, but are also receptive to other males. The explanation usually given for unfaithful female behavior is that most females are unable to bond with a male they would prefer as genetic father to their offspring. To secure male assistance the female therefore pairs with an available male but also copulates with males of supposedly higher genetic quality. Here we offer an alternative evolutionary explanation for female infidelity, which does not rely upon this 'Good Genes hypothesis of female choice. We show with a simple model that in an evolutionary game between three players, a male, a female and a male lover, solutions exist in which the female can secure more assistance from her mate by being receptive to other males. We conclude that female sexuality can have a decisive role in regulating social behaviour, in which the fertile female is the driving force.