Application of an immunosensor for the detection of the β-lactam antibiotic, cephalexin

Public concern surrounding antibiotic contamination in food and food products has made it imperative to develop analytical methods for their detection. Polyclonal antibodies were used in the development of a surface plasmon resonance (SPR)-based inhibition immunoassay for cephalexin. A conjugate consisting of cephalexin-bovine serum albumin (BSA) was immobilized on the dextran gel surface of the sensor chip. Binding/regeneration studies of antibody to immobilized cephalexin were studied and dissociation of the antibody from the immobilized cephalexin was easily achieved with 10 mmol l^-1 NaOH. Forty surface regeneration cycles were carried out and found to be reproducible with only a 7.4% decrease in binding over this number of regenerations. Model inhibition immunoassays for cephalexin were developed in PBS and spiked milk samples with detection ranges of 4.88 to 2,500 ng ml^-1 and 244 to 3,906 pg ml^-1, respectively.     

Preventive care: do we practice what we preach?

We used insurance claims from enrollees in the Rand Health Insurance Experiment to determine the amount of selected components of preventive care received by a representative sample of the non-aged population in the United States and to determine whether insurance coverage was an important determinant of that amount. Only 45 percent of infants received timely immunization for DPT and polio; 93 per cent received some well child care by 18 months of age. In the three-year experimental period, only 4 per cent of adults had a tetanus shot, 66 per cent of women aged 17-44 and 57 per cent aged 45-65 received a Pap smear, and 2 per cent of women aged 45-65 had a mammogram. Cost sharing was associated with even less preventive care: 60 per cent of children on the free plan and 49 per cent on cost sharing plans received preventive care of any type. For adults, women on the free plan received more preventive care of several kinds, and those aged 45-65 received more Pap smears than those on cost-sharing plans. Even with free care, most enrollees did not receive adequate preventive care. Thus, free care alone, while significant, is not a sufficient incentive to providing recommended levels of preventive care. The average per person insurance charge for increasing the amount of preventive care to a level consistent with that recommended would be $22 for a complete set of immunizations by age 18 months, $9 for a Pap smear every three years, and $97 for a Pap test and mammogram every three years.     

Solid-phase synthesis of 16 potent (selective and nonselective) in vivo antagonists of oxytocin

We describe the synthesis and some pharmacological properties of 16 new in vivo antagonists of oxytocin. These are based on modifications of three peptides: A, B, and C. A is our previously reported potent and selective antagonist of the vasopressor (V1 receptor) responses to arginine-vasopressin (AVP)/weak oxytocin antagonist, [1-(beta-mercapto-beta,beta-pentamethylenepropionic acid), 2-O-methyltyrosine]arginine-vasopressin (d(CH2)5[Tyr(Me)2]AVP. B reported here, the Ile3 analogue of A, is d(CH2)5[Tyr(Me)2]AVT (5 below) and C is our previously reported potent nonselective oxytocin antagonist/AVP V1 antagonist, [1-(beta-mercapto-beta,beta-pentamethylenepropionic acid),2-O- methyltyrosine,8-ornithine]vasotocin (d(CH2)5[Tyr(Me)2]OVT). The following substitutions and deletions, alone or in combination, were employed in A, B, and C: 1-deaminopenicillamine (dP); D-Tyr(Alk)2 (where Alk = Me or Et), D-Phe2; Val4, Thr4; delta 3-Pro7; Lys8, Cit8; desGly9, desGly-NH2(9), Ala-NH2(9); Leu-NH2(9); Arg-NH2(9). The 16 new analogues are (1) d(CH2)5[D-Tyr(Me)2]AVP, (2) d(CH2)5[D-Tyr(Me)2, Val4,delta 3-Pro7]AVP, (3) d(CH2)5[D-Tyr-(Et)2, Val4,Lys8]VP, (4) d(CH2)5[D-Tyr(Et)2,Val4,Cit8]VP, (5) d(CH2)5[Tyr(Me)2]AVT, (6) d(CH2)5[Tyr(Me)2,Lys8]VT, (7) dP[Tyr(Me)2]AVT, (8) dP[Tyr(Me)2,Val4]AVT, (9) d(CH2)5[D-Tyr(Me)2, Val4]AVT, (10) d(CH2)5[D-Phe2,Val4]AVT, (11) d(CH2)5[Tyr(Me)2,Thr4]OVT, (12) d(CH2)5[Tyr(Me)2,Thr4,Ala-NH2(9)]OVT, (13) d(CH2)5[Tyr(Me)2,Thr4,Leu-NH2(9)]OVT, (14) d(CH2)5[Tyr(Me)2,Thr4,Arg-NH2(9)]OVT, (15) desGly-NH2(9),d(CH2)5[Tyr(Me)2,Thr4]OVT, (16) desGly9,d(CH2)5[Tyr(Me)2,Thr4]OVT. 1-4 are analogues of A, 5-10 are analogues of B, and 11-16 are analogues of C. Their protected precursors were synthesized either entirely by the solid-phase method or by a combination of solid-phase and solution methods (1 + 8 or 8 + 1 couplings). All analogues were tested in rats for agonistic and antagonistic activities in oxytocic (in vitro, without and with Mg2+, and in vivo) assays as well as by antidiuretic and vasopressor assays. All analogues exhibit potent oxytocic antagonism in vitro and in vivo. With an in vitro pA2 (in the absence of Mg2+) = 9.12 +/- 0.09, dP[Tyr(Me)2]AVT is (7) one of the most potent in vitro oxytocin antagonists reported to date. Fifteen of these analogues (all but 6) appear as potent or more potent in vivo oxytocin antagonists than C (pA2 = 7.37 +/- 0.17). Analogues 1-9 and 14 are potent AVP V1 antagonists. Their anti-V1 pA2 values range from 7.92 to 8.45. They are thus nonselective oxytocin antagonists.(ABSTRACT TRUNCATED AT 400 WORDS)     

Considerations regarding the application of capillary optics to medical radiography.

The generation of monoenergetic or quasimonoenergetic x-ray spectra has been accomplished by several methods including the use of K-edge filtration, characteristic radiation, crystal monochromators, and multilayer mirrors. In this paper some practical questions are discussed regarding the possibility of obtaining narrow-band spectra using x-ray reflection in glass capillary optics which have been reported recently in connection with the focusing and generation of parallel x-ray beams. Derivation of formulas for double and triple reflection with and without additional K-edge filtration imply that these methods are superior to the use of filtration alone. It is shown that the double reflection scheme is most sensitive to the angular divergence of the beam emanating from the capillary array used to generate the incident parallel beam. Simulations using three reflections predict output spectra which are relatively insensitive to blurring due to angular divergence. A small amount of K-edge filtration in combination with three reflections provides considerable sharpening of the x-ray spectrum. Aside from the spectral selectivity provided by the capillary array, the reduced divergence of the primary transmission may have advantages for scatter rejection in situations where the air gap between the patient and detector is increased by an amount consistent with the resolution requirements of the application. In parallel beam situations, the use of capillary arrays between the patient and detector may also be useful for the rejection of divergent scattered radiation.     

Pneumothorax and other lung diseases: effect of altered resolution and edge enhancement on diagnosis with digitized radiographs

Prior studies have shown that pneumothorax is one of the more difficult entities to diagnose with digitized radiography. This study was designed to test whether increasing resolution from 1.25 to 2.5 line pairs per millimeter (lp/mm) and image processing (edge enhancement from unsharp masking) would increase accuracy and confidence in the diagnosis of pneumothorax, as well as normal cases and other forms of lung disease. Conventional radiographs were digitized with use of a laser reader and then reformatted as film hard copy. Eleven observers read 35 cases reformatted in three different ways (1.25 lp/mm, 2.5 lp/mm, 1.25 lp/mm unsharp mask). The images with finer resolution (2.5 lp/mm) and unsharp mask images were superior to those with coarser resolution (1.25 lp/mm) for the diagnosis of pneumothorax. There was no difference in diagnostic accuracy for normal patients. For abnormalities other than pneumothorax, the unsharp mask images were significantly worse. Confidence in the diagnosis of pneumothorax and other abnormalities was highest with the finest resolution (2.5 lp/mm).     

Paraimmunoblastic variant of small lymphocytic lymphoma/leukemia

We report 16 cases of a distinctive, biologically aggressive variant of small lymphocytic lymphoma/leukemia (SLL/L) that is characterized by the diffuse proliferation of cells normally comprising the pseudoproliferation centers (so-called paraimmunoblasts). Demographically, the patients differed in no significant regard from patients with SLL/L of usual type. Rapidly progressive, generalized lymphadenopathy was the dominant clinical finding in 15 of the 16 patients; one patient presented with symptoms related to lymphomatous involvement of the stomach and regional lymph nodes. Splenomegaly was observed in five patients. Seven patients, two of whom had a history of indolent-phase chronic lymphocytic leukemia, had an absolute lymphocytosis at diagnosis. In most patients, bone marrow involvement was noted at diagnosis. It consisted predominantly of small lymphocytic infiltrates indistinguishable from those observed in SLL/L of usual type; significant paraimmunoblastic infiltration was infrequent and generally occurred late in the disease course. Immunohistochemical and cytogenetic study further substantiated the hypothesized relationship of these cases to SLL/L. Findings included (a) coexpression of sIg and Leu-1 antigen in the majority of cases and (b) the presence of a t(11;14) (q13;q32) chromosome translocation in two of three cases with analyzable metaphases. Although treatment protocols were not uniform, follow-up data indicated an accelerated clinical course. Eleven patients have died of their disease between 3 and 39 months after diagnosis; the median survival was 28 months.     

Assessment of intracardiac shunts with gadolinium-enhanced ultrafast MR imaging.

Dye dilutional techniques are widely accepted for the assessment of intracardiac shunts, but current techniques require arterial access or radioisotope injection. Ultrafast (less than 500 msec) magnetic resonance (MR) imaging is ideally suited for the evaluation of an indicator during passage through the heart. Twenty patients were studied, including 13 with shunts. Four-chamber, T1-weighted images were obtained during bolus injection of gadopentetate dimeglumine. A single image was obtained in 420 msec, with repetitive images acquired after each QRS complex. After the contrast material was injected, there was pronounced signal intensity enhancement in the right atrium, followed by the right ventricular cavity, left atrium, left ventricular cavity, and descending aorta. Patients with substantial intracardiac shunts demonstrated early recirculation. First-pass contrast material-enhanced MR imaging is a promising new technique for the rapid assessment of intracardiac shunts. Combined with anatomic and functional MR imaging techniques, it can help provide a comprehensive noninvasive evaluation of suspected intracardiac shunts or provide follow-up in patients with known shunts.     

Incidence of otitis media in vitamin A-deficient guinea pigs.

After observing an association between vitamin A deficiency and otitis media among children in Micronesia, we sought to develop an animal model for vitamin A-deficiency-induced otitis media. Thirty juvenile guinea pigs were screened for absence for otitis and then divided into two groups. The experimental group was placed on a vitamin A-free diet, whereas control animals were fed a standard formula. The animals were followed by means of otologic examination and serum retinol determinations. When experimental animals demonstrated vitamin A deficiency, they were killed along with one control and temporal bones were harvested and prepared for histologic examination. None of the 15 controls demonstrated middle ear abnormalities. In the experimental group, 77% of temporal bones showed middle ear pathology consisting of either of subepithelial edema (27%) or frank otitis media (50%). These results indicate that the guinea pig is a suitable species for study of otitis media in association with vitamin A deficiency. We postulate that eustachian tube dysfunction is the primary mechanism for inducing middle ear inflammation in this animal model.     

Digit ratio and hand-grip strength in German and Mizos men: cross-cultural evidence for an organizing effect of prenatal testosterone on strength.

The ratio of the length of the second and fourth digits (2D:4D), a putative proxy of prenatal testosterone (PT), is correlated with measures of physical fitness. The relationship between the organizing effect of PT on physical fitness is likely to arise as a response to intrasexual male competition for females. Physical strength is also likely to be important in intrasexual conflict, but nothing is known concerning the relationship between 2D:4D and strength. The strength of an individual is strongly influenced by body size, and 2D:4D is strongly dependent on ethnicity. We present evidence that strength, as measured from hand-grip strength, is related to 2D:4D in samples from two ethnic groups (52 Caucasian men from Germany, and 88 Oriental Mizos men from northeast India) which differed markedly in size. We found that 1) the German men were heavier and stronger, but had higher 2D:4D (lower PT) than the Mizos men; 2) a median split for grip strength into low (LGS) and high (HGS) groups showed that for right-hand 2D:4D (but not left-hand 2D:4D), the LGS men had higher 2D:4D than the HGS men; and 3) the relationships between right 2D:4D and grip strength were independent of ethnicity, age, height, and weight. Measures of grip strength correlate strongly with strength in other muscle groups, so we conclude that PT may have an early organizing effect on strength in men, and this is likely to be widespread in human groups.