Formulary review of the carbapenems: comparison of imipenem/cilastatin and meropenem

The duplicative mutation of an Ala-Val-Arg sequence at positions 208 to 210 in the loop structure of Enterobacter cloacae class C beta-lactamase caused substrate specificity extension to oxyimino beta-lactam antibiotics and this chromosomal mutation provided bacterial cells with high resistance to the beta-lactams (M. Nukaga et al, 1995, J. Biol. Chem. 270, 5729-5735). In order to confirm the universality of this phenomenon among other class C beta-lactamases, the duplicative mutation was applied to a class C beta-lactamase of Citrobacter freundii, which has 74% homology to the E. cloacae beta-lactamase amino acid sequence. The counterpart sequence to the Ala-Val-Arg of the E. cloacae enzyme in C. freundii beta-lactamase was identified to be Pro-Val-His. A Pro-Val-His sequence was inserted just after the native Pro-Val-His sequence at positions 208 to 210 in the C. freundii beta-lactamase. The resulting mutant of C. freundii beta-lactamase obtained a striking characteristic that we expected, showing substrate specificity extension to oxyimino beta-lactams. Nearly the same result was obtained with the insertion of an Ala-Val-Arg sequence after the native Pro-Val-His sequence. These results indicate that structural modification of this locus commonly induces modification of the substrate specificity to unfavorable substrates for many chromosomal class C beta-lactamases produced by gram-negative bacteria.     

Protein kinase C in platelets of depressed patients

Though described in 1769, the etiology of Zenker's diverticulum remains unclear. Various primary esophageal motor disorders have been proposed, but no consistent manometric pattern or anatomic etiology has been uniformly recognized. An association with clinical neurologic disease at our institution prompted a review of 12 cases of Zenker's diverticulum in patients over 60 years of age, treated in the last 8 years. Nine patients (75%) underwent cricopharyngeus myotomy and diverticulectomy, with uniformly good results. Ten patients (83%) had an associated neurologic disorder, substantiated by cranial CT or MRI, in most cases. A wide range of neurologic problems were identified, but a strong trend toward brainstem or basilar lesions was present. As expected, the etiology of the neurologic abnormality in most patients in this group was cerebrovascular disease, but two patients had peripheral neuropathies. We suggest that the etiology of Zenker's diverticulum in the elderly may be neurologic in origin. Esophageal motor disorders, including incomplete upper esophageal sphincter opening and increased hypopharyngeal pressures, which may result in Zenker's diverticulum, may be a manifestation of central or peripheral neurologic disease in the elderly.     

Hormone replacement therapy and risk of hip fracture: population based case-control study. The Swedish Hip Fracture Study Group

OBJECTIVE: To determine the relative risk of hip fracture associated with postmenopausal hormone replacement therapy including the effect of duration and recency of treatment, the addition of progestins, route of administration, and dose. DESIGN: Population based case-control study. Setting: Six counties in Sweden. SUBJECTS: 1327 women aged 50-81 years with hip fracture and 3262 randomly selected controls. MAIN OUTCOME MEASURE: Use of hormone replacement therapy. RESULTS: Compared with women who had never used hormone replacement therapy, current users had an odds ratio of 0.35 (95 % confidence interval 0.24 to 0.53) for hip fracture and former users had an odds ratio of 0.76 (0.57 to 1.01). For every year of therapy, the overall risk decreased by 6% (3% to 9%): 4% (1% to 8%) for regimens without progestin and 11% (6% to 16%) for those with progestin. Last use between one and five years previously, with a duration of use more than five years, was associated with an odds ratio of 0.27 (0.08 to 0.94). After five years without hormone replacement therapy the protective effect was substantially diminished (-7% to 48%). With current use, an initiation of therapy nine or more years after the menopause gave equally strong reduction in risk for hip fracture as an earlier start. Oestrogen treatment with skin patches gave similar risk estimates as oral regimens. CONCLUSIONS: Recent use of hormone replacement therapy is required for optimum fracture protection, but therapy can be started several years after the menopause. The protective effect increases with duration of use, and an oestrogen-sparing effect is achieved when progestins are included in the regimen.     

The effect of recombinant human erythropoietin on hemostasis, fibrinolysis, and blood rheology in autologous blood donors

We report three cases of Castleman's disease mimicking the features of collagen disease. Case 1: A 39-year-old woman presented with intermittent arthralgia and fever. Laboratory findings were positive results for antinuclear antibody (80x speckled type), the LE test, anti-SSA antibody, anti-RNP antibody, and Coombs test. The patient was suspected to have systemic lupus erythematosus (SLE) or Sj?gren syndrome, but a lymph node biopsy revealed the plasma cell type of Castleman's disease. Steroid treatment led to resolution of her symptoms. Case 2: A 60-year-old man with mixed type Castleman's disease had proteinuria with renal dysfunction, autoimmune thrombocytopenia, antinuclear antibody, anti-RNP antibody, anti-DNA antibody and anti-cardiolipin antibody. The patient was suspected to have SLE but cervical lymph node biopsy revealed the mixed type of Castleman's disease. Symptoms were not controlled with steroid therapy. He developed renal failure that required for hemodialysis and died of gastrointestinal bleeding due to severe thrombocytopenia. Case 3: A 46-year-old woman had Raynaud's phenomenon, sclerodactylia, and nail fold bleeding. Laboratory tests were revealed positive for antinuclear antibody, anti-ENA antibody, and LE cell preparation. Radiographic study showed multiple masses in the retroperitoneal spaces, which necessitated laparotomy. Firstly, the patient was suspected to have systemic sclerosis or mixed connective tissue disease (MCTD). A biopsy revealed the hyaline-vascular type of Castleman's disease. The serum level of IL-6 by ELISA was high in all of three cases. In case 1, symptoms improved and the IL-6 level normalized after steroid treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

CVT-313, a specific and potent inhibitor of CDK2 that prevents neointimal proliferation

The activity of cyclin-dependent kinase 2 (CDK2) is essential for progression of cells from G1 to the S phase of the mammalian cell cycle. CVT-313 is a potent CDK2 inhibitor, which was identified from a purine analog library with an IC50 of 0.5 microM in vitro. Inhibition was competitive with respect to ATP (Ki = 95 nM), and selective CVT-313 had no effect on other, nonrelated ATP-dependent serine/threonine kinases. When added to CDK1 or CDK4, a 8.5- and 430-fold higher concentration of CVT-313 was required for half-maximal inhibition of the enzyme activity. In cells exposed to CVT-313, hyperphosphorylation of the retinoblastoma gene product was inhibited, and progression through the cell cycle was arrested at the G1/S boundary. The growth of mouse, rat, and human cells in culture was also inhibited by CVT-313 with the IC50 for growth arrest ranging from 1.25 to 20 microM. To evaluate the effects of CVT-313 in vivo, we tested this agent in a rat carotid artery model of restenosis. A brief intraluminal exposure of CVT-313 to a denuded rat carotid artery resulted in more than 80% inhibition of neointima formation. These observations suggest that CVT-313 is a promising candidate for evaluation in other disease models related to aberrant cell proliferation.     

The isolability of the causative agent of pseudotuberculosis from rodents in Georgia

Since 1960 the cases of isolating the pseudopathogen agent have been recorded in Georgia at particular regular intervals. The accumulated material mainly involves epizootiological findings, which may provide a definite insight into the prevalence of this disease in Georgia. In 1960-1990, 31 strains were isolated in particular regions and populated areas; these included 17 strains from grey rats, 4 from house mice, 9 from common voles, and 1 from field mice. The particular constancy of isolation of the pseudotuberculosis pathogen from synanthropic rodents indicates that they may really infect humans, contrary to the opinion of G. P. Somov et al. who consider that among the synanthropic rodents there is only a chronic epizootic causing no animal death and that the role of the rodents in the spread of infection among humans is insignificant. However, doubt is cast on the validity of this proposition as the evidence for carriage alone among the rodents is lacking. The special literature contains no information on the dynamics of an epizootic process of pseudotuberculosis among the synanthropic rodents whereas the high pathogen detection rate in the viscera of these animals, recognized by Somov et al., mostly likely confirms the possibility of the course of epizootic manifested by the disease, but not only carriage. There is reason to consider pseudotuberculosis to be a naturally focal disease that is characterized by recurrent epizootics of varying intensity. Therefore, wild and synanthropic rodents may be as a reservoir and a source of infection. The circulation of the pathogen among wild and synanthropic rodents, in terms of their ability to be preserved and breed in the environmental objects, presents a permanent risk of not only sporadic cases, but also pseudotuberculosis outbreaks.     

The effects of amount of whole barley, barley bulk density, and form of roughage on feedlot lamb performance, carcass characteristics, and digesta kinetics

We conducted two feedlot trials and one metabolism trial to evaluate the effect of barley level, barley bulk density, and physical form of roughage on lamb growth performance and digesta kinetics. Level of whole barley (50, 70, 90%) and type of roughage (chopped or pelleted alfalfa) were evaluated in Trial 1 (50 d period). Trial 2 (50 d) evaluated barley bulk density (heavy = 671 and light = 607 kg/m3), form of roughage (pelleted or chopped alfalfa), and level of barley (80 or 40%). The influence of treatments used in Trial 2 on digesta kinetics was evaluated in Trial 3. Gain:feed increased and DMI decreased (P < .10) linearly with increasing level of barley, and ADG and DMI were greater (P < . 10) for lambs fed pelleted vs chopped alfalfa in Trial 1. The 70% barley diet produced the highest yield grade and kidney-pelvic fat and the lowest leg score among barley levels (P < .10). Lambs fed pelleted alfalfa had heavier carcasses and a thicker body wall than lambs fed chopped alfalfa (P < .02). In Trial 2, DMI was less and gain:feed greater (P < .01) for lambs fed the heavy barley than for lambs fed the light barley and for the 80% barley diet compared to the 40% barley diet. Lambs fed pelleted alfalfa had greater dressing percentages than lambs fed chopped alfalfa. Backfat and body wall thickness were greater (P < .10) for lambs fed the 80% barley diet than for those fed the 40% barley diet. In Trial 3, retention time of barley was greater (P < .10) for lambs fed light rather than heavy barley, and retention time of alfalfa was greater (P < .10) for lambs fed chopped compared with pelleted alfalfa. Acetate:propionate ratio was greater (P < .10) for lambs fed light vs heavy barley and lambs fed the 40 vs 80% barley diets. Ruminal pH was lower (P = .05) and in situ barley digestion greater (P = .03) over time in lambs fed the 80% barley diet than in lambs fed the 40% barley diet. Feedlot lamb ADG was not always greatest with high levels of barley; however, gain:feed improved at the higher barley levels. The higher barley levels seemed to result in fatter lambs.     

Mechanism of Ca2+ and monosaccharide binding to a C-type carbohydrate-recognition domain of the macrophage mannose receptor

Site-directed mutagenesis has been used to identify residues that ligate Ca2+ and sugar to the fourth C-type carbohydrate-recognition domain (CRD) of the macrophage mannose receptor. CRD-4 is the only one of the eight CRDs of the mannose receptor to exhibit detectable monosaccharide binding when expressed in isolation, and it is central to ligand binding by the receptor. CRD-4 requires two Ca2+ for sugar binding, like the CRD of rat serum mannose-binding protein (MBP-A). Sequence comparisons between the two CRDs suggest that the binding site for one Ca2+, which ligates directly to the bound sugar in MBP-A, is conserved in CRD-4 but that the auxiliary Ca2+ binding site is not. Mutation of the four residues at positions in CRD-4 equivalent to the auxiliary Ca2+ binding site in MBP-A indicates that only one, Asn728, is involved in ligation of Ca2+. Alanine-scanning mutagenesis was used to identify two other asparagine residues and one glutamic acid residue that are probably involved in ligation of the auxiliary Ca2+ to CRD-4. Sequence comparisons with other C-type CRDs suggest that the proposed binding site for the auxiliary Ca2+ in CRD-4 of the mannose receptor is unique. Evidence that the conserved Ca2+ in CRD-4 bridges between the protein and bound sugar in a manner analogous to MBP-A was obtained by mutation of one of the amino acid side chains at this site. Ring current shifts seen in the 1H NMR spectra of methyl glycosides of mannose, GlcNAc, and fucose in the presence of CRD-4 and site-directed mutagenesis indicate that a stacking interaction with Tyr729 is also involved in binding of sugars to CRD-4. This interaction contributes about 25% of the total free energy of binding to mannose. C-5 and C-6 of mannose interact with Tyr729, whereas C-2 of GlcNAc is closest to this residue, indicating that these two sugars bind to CRD-4 in opposite orientations. Sequence comparisons with other mannose/GlcNAc-specific C-type CRDs suggest that use of a stacking interaction in the binding of these sugars is probably unique to CRD-4 of the mannose receptor.     

The fetoplacental pressor effects of low-dose acetylsalicylic acid and angiotensin II in the ex vivo cotyledon model

OBJECTIVE: Our purpose was to investigate perfusion pressure changes ex vivo induced by angiotensin II on fetoplacental vasculature pretreated with low-dose acetylsalicylic acid. STUDY DESIGN: Two cotyledons from each of 12 placentas were perfused. The intervillous space of one cotyledon was infused with acetylsalicylic acid (5 x 10(-5) mol/L) similar to the serum concentration of women receiving daily low-dose aspirin therapy (60 to 81 mg). The control cotyledon was infused with an equivalent amount of normal saline solution. Two doses of angiotensin II, 1 x 10(-11.5) and 1 x 10(-10) moles, were injected as boluses into the chorionic arteries of each cotyledon. A 3 x 10(-7) mole dose of angiotensin II was also injected into the intervillous space. Statistical analysis was performed with analysis of variance, and results are expressed as mean pressure change in millimeters of mercury +/- SEM. RESULTS: Perfusion pressure response did not vary between cotyledons pretreated with acetylsalicylic acid and control cotyledons when 3 x 10(-7) moles of angiotensin II was injected into the intervillous space (8.0 +/- 1.9 mm Hg vs 9.8 +/- 1.6 mm Hg, p = 0.59). There were no differences between cotyledons in pressure response to 1 x 10(-11.5) moles of angiotensin II injected into the fetal circuit (5.9 +/- 0.8 mm Hg vs 6.7 +/- 0.9 mm Hg, p = 0.51). However, in the cotyledons pretreated with acetylsalicylic acid there was a decrease in the pressor response to 1 x 10(-10) moles of angiotensin II (14.1 +/- 1.4 mm Hg vs 21.5 +/- 3.3 mm Hg, p = 0.05). CONCLUSIONS: Low-dose aspirin infused into the intervillous space decreases vasoconstriction elicited by angiotensin II in the fetoplacental compartment. This suggests that maternal low-dose aspirin therapy has effects in the fetoplacental circulation in addition to its effects in the maternal circulation.     

Selective inhibition of T cell proliferation but not expression of effector function by human alveolar macrophages

BACKGROUND: Alveolar macrophages are thought to play an important part in regulating lung immune responses. While it is clear that human alveolar macrophages suppress T cell proliferation in vitro, the mechanisms by which this is achieved are not clear, nor is it known whether alveolar macrophages also inhibit other aspects of T cell function. METHODS: Peripheral blood mononuclear cells were stimulated with phytohaemagglutinin or house dust mite allergen, and cultured with variable numbers of autologous alveolar macrophages obtained by bronchoalveolar lavage from 20 normal subjects. RESULTS: Alveolar macrophages induced a reversible inhibition of T cell proliferation in response to both mitogen and allergen stimulation, with the latter being considerably more susceptible to inhibition. This was achieved via heterogenous mechanisms, involving both soluble factors derived from alveolar macrophages and cell-cell contact. Despite inhibiting proliferation, alveolar macrophages had little or no effect on T cell calcium flux, the characteristic changes in CD3, CD2, CD28 and interleukin-2 (IL-2) receptor expression which accompany normal T cell activation, and IL-2 and interferon gamma secretion. In contrast, alveolar macrophages inhibited the tyrosine phosphorylation of proteins which may be involved in IL-2 receptor-associated signal transduction. CONCLUSIONS: The immunoregulatory properties of alveolar macrophages are relatively selective, allowing T cell activation and cytokine secretion while inhibiting T cell proliferation within the lung.