Fast cavity-enhanced atom detection with low noise and high fidelity.

Cavity quantum electrodynamics describes the fundamental interactions between light and matter, and how they can be controlled by shaping the local environment. For example, optical microcavities allow high-efficiency detection and manipulation of single atoms. In this regime, fluctuations of atom number are on the order of the mean number, which can lead to signal fluctuations in excess of the noise on the incident probe field. Here we demonstrate, however, that nonlinearities and multi-atom statistics can together serve to suppress the effects of atomic fluctuations when making local density measurements on clouds of cold atoms. We measure atom densities below 1 per cavity mode volume near the photon shot-noise limit. This is in direct contrast to previous experiments where fluctuations in atom number contribute significantly to the noise. Atom detection is shown to be fast and efficient, reaching fidelities in excess of 97% after 10 µs and 99.9% after 30 µs.

Immunohistochemistry to identify Leishmania parasites in fixed tissues.

The definitive diagnosis of leishmaniasis currently depends on the identification of characteristic amastigote morphology in tissue, or isolation of promastigotes by culture. Histopathological identification can be difficult, and is variably sensitive; culture is considered "the gold standard", but is not uniformly diagnostic or available. In this study, we compared light microscopic immunohistochemistry (IHC) using a monoclonal anti-Leishmania antibody (G2D10) to standard hematoxylin and eosin (H&E) stain in the diagnosis of Leishmania on skin. Sixty-one archived specimens from patients suspected of being infected with Leishmania were used; 41 of these had leishmaniasis confirmed by culture. Although not statistically significant, both sensitivity and specificity were higher for IHC compared to H&E: 51% (95% CI: 35-67%) compared to 42% (CI: 26-58%; 2p=0.29) for sensitivity, and 100% (CI: 83-100%) compared to 85% (CI: 62-97%, 2p=0.25) for specificity, respectively. Furthermore, because organisms could be diagnosed by IHC at low power (x20-40), this assay was more rapid than H&E, in which parasite morphology could best be identified at oil immersion power. The G2D10 antibody has broad Leishmania species recognition, and offers promise as a simple, rapid diagnostic screen for leishmaniasis. Further study is underway to better characterize this antibody.

The United States military and leishmaniasis.

Leishmaniasis occurs not only in American travelers and military personnel alike but infects a significant portion of the world's population. The US military has made major contributions to the understanding of the complicated epidemiology of this parasite, the development of rapid reliable diagnostic tests, and to the development of safe, more efficient, and more effective treatment of leishmaniasis.

Educational initiatives in long-term care--Midwest Bioethics Center's Kansas nursing home project.

Most Americans would doubtless agree that positive change is a critical need in facilities providing long-term care. This article describes a project currently underway in which Midwest Bioethics Center (MBC) staff and Kansas City area experts in long-term care are doing their part to create meaningful change in the way residents of nursing homes are cared for in the last chapter of their lives.

Acinic cell carcinoma presenting as an upper lip mass.

BACKGROUND: Acinic cell carcinoma (ACC) is a rare tumor generally involving the parotid gland and infrequently the minor salivary glands. OBJECTIVE: Clinical and histopathological characteristics of a case of ACC of the minor salivary glands, and brief review of the literature. METHODS: Routine stains with H&E, PAS, iron, and mucicarmine; immunohistochemistry with cytokeratin, CEA, S-100, and leu-M1. RESULTS: The ACC was removed by wide local excision. Histopathology revealed papillary cystic, solid, and ductal architectural features. The mucinous material within the lumina of the cystic and ductal structures was PAS and mucicarmine positive, and diastase resistant. All tested immunohistochemical reagents reacted with the epithelial cells. There were no mitoses, necrosis, cellular pleomorphism, or infiltration. CONCLUSIONS: ACC is a rare tumor of salivary glands characterized by an indolent clinical course with the potential for both local recurrence and metastatic spread when tracked for decades. Published mortality rates vary from 6 to 50%. Histopathologic features do not reliably predict biologic behavior.

Nitric oxide production during Vibrio cholerae infection.

Vibrio cholerae induces massive intestinal fluid secretion that continues for the life of the stimulated epithelial cells. Enhanced regional blood flow and peristalsis are required to adapt to this obligatory intestinal secretory challenge. Nitric oxide (NO) is a multifunctional molecule that modulates blood flow and peristalsis and possesses both cytotoxic and antibacterial activity. We demonstrate that, compared with those in asymptomatic control subjects, levels of stable NO metabolites (NO2-/NO3-) are significantly increased in sera from acutely ill Peruvian patients with natural cholera infection as well as from symptomatic volunteers from the United States infected experimentally with V. cholerae. In a rabbit ileal loop model in vivo, cholera toxin (CT) elicited fluid secretion and dose-dependent increases in levels of NO2-/NO3- in the fluid (P < 0.01). In contrast, lipopolysaccharide (LPS) elicited no such effects when applied to the intact mucosa. NO synthase (NOS) catalytic activity also increased in toxin-exposed tissues (P < 0.05), predominantly in epithelial cells. The CT-induced NOS activity was Ca2+ dependent and was not suppressed by dexamethasone. In conclusion, symptomatic V. cholerae infection induces NO production in humans. In the related animal model, CT, but not LPS, stimulated significant production of NO in association with increases in local Ca(2+)-dependent NOS activity in the tissues.

Delayed-type hypersensitivity skin testing using third variable loop peptides identifies T lymphocyte epitopes in human immunodeficiency virus-infected persons.

Cellular immune responses to human immunodeficiency virus type 1 (HIV-1) infection, particularly in vivo responses, have been difficult to study in large patient cohorts because of technical impediments. By use of small peptide fragments of the HIV-1 gp120 third variable loop, the CD4 T lymphocyte epitopes of 2 HIV-infected persons were mapped using a cutaneous delayed-type hypersensitivity (DTH) assay. The in vivo DTH responses correlated with epitopes previously identified in vitro using CD4 T lymphocyte lines. The ability to determine CD4 T lymphocyte epitopes in large cohorts of patients using this simple in vivo technique would provide important diagnostic and prognostic data regarding effective immunoregulation of HIV-1. This technique should have broad applicability in HIV vaccine development and in the investigation of other immune-mediated human diseases.

Peru-15, an improved live attenuated oral vaccine candidate for Vibrio cholerae O1.

Cholera vaccine candidate Peru-15 was derived from a Vibrio cholerae O1 El Tor Inaba strain by deleting the cholera toxin genetic element, introducing the gene encoding cholera toxin B subunit into recA, and screening for nonmotility. In a controlled study, Peru-15 (2 x 10(8) cfu) was administered to 11 volunteers. No vaccinee developed diarrhea, and 10 of 11 had > 4-fold rises in vibriocidal antibody titers. One month later, 5 vaccinees and 5 control volunteers were challenged with wild type V. cholerae O1. Four of 5 controls developed diarrhea (mean, 1.9 L). Two Peru-15 vaccinees developed diarrhea, 1 with < 0.3 L and 1 with approximately 1.0 L; this latter volunteer had not developed a significant vibriocidal immune response to vaccination. Peru-15 shows promise as a single-dose, oral cholera vaccine that is safe, immunogenic, and protective.

Safety, immunogenicity, and efficacy of live attenuated Vibrio cholerae O139 vaccine prototype.

New vaccines are needed to prevent cholera caused by Vibrio cholerae O139. Attenuated V cholerae O139 vaccines were made by deleting multiple copies of the cholera-toxin genetic element from two virulent strains of the organism, MO10 and AI4456. The deletion mutants were further modified by insertion of a construct that encoded the B subunit of cholera toxin, thus generating strains Bengal-3 and VRI-16. A stable spontaneous non-motile derivative of Bengal-3 was isolated and designated Bengal-15; VRI-16 is naturally non-motile. Bengal-3, Bengal-15, and VRI-16 were evaluated as oral single-dose cholera vaccine candidates in 4 volunteers each, and MO10 was given to 3 volunteers. 1 of 4 volunteers who received Bengal-3 and all 3 who received MO10 had diarrhoea. VRI-16 caused no significant symptoms but was not immunogenic. Bengal-15 produced few symptoms and was nearly as immunogenic as MO10. Subsequently, Bengal-15 was given to 10 volunteers at a dose of 10(8) colony-forming units. No volunteers had diarrhoea, and other subjective symptoms were as common in vaccinees as in 3 buffer recipients. 1 month after vaccination, 7 vaccinees, the 3 buffer recipients, and 3 unimmunised subjects were challenged with 5 x 10(6) colony-forming units of V cholerae O139. 5 of 6 controls had cholera-like diarrhoea. By contrast, 1 of 7 vaccinees had diarrhoea, which was mild and had a long incubation period. Vaccine protective efficacy was 83%. Our results indicate the Bengal-15 is a safe live attenuated vaccine candidate for cholera caused by the O139 serogroup.

Development of a live, oral, attenuated vaccine against El Tor cholera.

Vibrio cholerae El Tor strains from Peru, Bangladesh, and Bahrain were attenuated by deletion of a genetic element that encodes virulence factors and RS1. The B subunit of ctx (ctxB) was reintroduced into the recA gene of the deletion mutants, rendering them unable to recombine with exogenous genetic elements and generating Peru-3, Bang-3, and Bah-3. Fifteen volunteers received one dose of various vaccine strains at 4 x 10(6) to 1 x 10(8) cfu. All strains colonized the gut. A > or = 4-fold rise in vibriocidal titer was observed in 14 volunteers, with titers of > or = 1600 in 13. Peru-3 was the least reactogenic, but 2 of 6 volunteers had loose stools. Peru-14, a filamentous motility-deficient mutant of Peru-3, was well tolerated and colonized 18 of 21 volunteers at doses of 2 x 10(6) to 1 x 10(9) cfu. Also, when 8 Peru-3 or Peru-5 vaccinees, 5 Peru-14 vaccinees, and 8 controls were challenged with 2 x 10(6) cfu V. cholerae El Tor Inaba (N16961), 11 vaccinees were protected compared with no controls. Peru-14 shows promise as a safe, effective, single-dose oral vaccine against El Tor cholera.

The effect of bromocryptine-induced hypoprolactinemia on xenogeneic and allogeneic skin graft survival in a mouse model.

Bromocryptine-induced hypoprolactinemia produces immunosuppression capable of extending allograft survival in solid organ transplantation models, but its potential for delaying either xenogeneic or allogeneic skin graft rejection is as yet undefined. In this study the ability of bromocryptine to prolong the survival of either cutaneous human xenografts or murine allografts was compared with cyclosporin A in a mouse model. In xenograft experiments cryopreserved human cadaver skin was grafted to B6D2F1 mice. In allograft experiments skin graft donors (A/J mice) differed genetically from recipients (Balb-C mice) at the major histocompatibility locus. In the untreated control group xenograft survival averaged 7 days and allograft survival 8 days. Xenograft rejection was delayed significantly and nearly equally by both bromocryptine and cyclosporin A (mean 15 and 16 days, respectively). However, allograft rejection was delayed longer by cyclosporin A (mean 24 days; p = 0.0002) than by bromocryptine (mean 15 days; p = 0.066). Administration of porcine prolactin (1 mg) partially reversed the bromocryptine effect (mean xenograft survival, 11 days; mean allograft survival, 10 days). The results of this study suggest that the immunosuppression of prolactin release by bromocryptine may be an effective, benign means of prolonging the survival of biologic dressings (e.g., banked cadaver allograft or porcine xenograft) used for temporary wound coverage in massively burned patients.

Role of adrenal cortical activation in the immunosuppressive effects of chronic morphine treatment.

Implantation of a 75-mg morphine pellet in sham-adrenalectomized male C3H/HeN mice resulted in significant elevations of serum corticosterone levels within 6 h. Corticosterone levels remained elevated (3- to 4-fold) for 72 h and had returned to normal by 120 h postimplantation. Within 48 h of pellet implantation, morphine-pelleted mice exhibited marked reductions in spleen (35%) and thymus weight (56%) relative to values in placebo-pelleted controls. In addition, adrenal hypertrophy was observed in the morphine-pelleted shams (50% increase in adrenal weight relative to placebo. The magnitude of splenic and thymic atrophy was reduced by about 50% in adrenalectomized morphine-pelleted mice (17% and 22% reductions, respectively) compared to that in adrenalectomized mice implanted with placebo pellets. Lymphocyte proliferative responses to the T-cell mitogen Concanavalin-A and the B-cell mitogen bacterial lipopolysaccharide were also significantly reduced in the morphine-pelleted sham mice. Morphine-induced suppression of Concanavalin-A- or lipopolysaccharide-stimulated lymphocyte proliferation was absent in adrenalectomized mice. Effects similar to adrenalectomy (e.g. lessening of magnitude of morphine-induced suppression of lymphoid organ weight and lymphocyte proliferation) were found in morphine-pelleted mice given the glucocorticoid receptor antagonist RU-486 at a dose of 10 mg/kg, twice daily. These studies imply that morphine-induced immunosuppression is at least in part mediated by the increase in serum corticosterone levels after implantation of the morphine pellet.

Cardiovascular effects of dynorphin A (1-13) in conscious rats and its modulation of morphine bradycardia over time.

The short-term cardiovascular effects of dynorphin A (1-13), as well as its effects upon morphine bradycardia were investigated. In unanesthetized, unrestrained rats, intracerebroventricular (ICV) dynorphin A (1-13) injections (10-20 micrograms) produced a dose-related pressor effect, whereas intravenous (IV) dynorphin A (1-13) (1.0 mg/kg) produced a depressor effect; these responses persisted less than five min. Heart rate was not significantly altered by these doses or routes of administration. Dynorphin A (1-13) also produced behavioral effects in the unanesthetized animals, such as wet dog shakes in response to IV administration and wet dog shakes accompanied by barrel rolling in response to ICV administration. To evaluate the effects of dynorphin A (1-13) pretreatment on the bradycardic response to IV morphine, rats were pretreated with 10 micrograms dynorphin A (1-13) ICV four, six or eight hours prior to challenge with morphine sulfate (0.1 mg/kg IV). Four hour pretreatment with dynorphin A (1-13) (tested at 14:00 hr) resulted in a potention of morphine bradycardia, with six hours pretreatment (tested at 16:00 hr) no effect was observed, and eight hours following dynorphin A (1-13) pretreatment (tested at 18:00 hr) morphine bradycardia was attenuated. Additionally, the bradycardic response to IV morphine alone became more exaggerated as rats approached their nocturnal activity cycle. These data further establish that dynorphin A (1-13) exerts a potent, long lasting modulatory effect on morphine bradycardia and emphasize the importance of circadian variables in altering the magnitude of cardiovascular responses to opioid agonists.

Repeated electroconvulsive shock: effect on sodium dependency and regional distribution of opioid-binding sites.

The effects of single and repeated electroconvulsive shock (ECS) on the binding of [3H]diprenorphine to rat brain membranes was studied. Repeated but not single ECS significantly increased the Bmax of [3H]diprenorphine binding when measured in the absence but not in the presence of NaCl. On a regional basis the effect of ECS was greatest in the olfactory bulb, nucleus accumbens, and striatum. More modest increases were found in the hippocampus, amygdala, septum, hypothalamus, and pyriform cortex. No significant effect was found in the brainstem and frontal cortex. Although the regional rank order of receptor increase does not match the receptor distribution of brain enkephalins, the receptor increase does parallel the regional increases in brain enkephalins following ECS.

Evidence for delta-receptor involvement in the post-ictal antinociceptive responses to electroconvulsive shock in rats.

Studies were conducted with the putative delta-opioid receptor antagonist lCl 154,129 (lCl) and the putative mu-receptor antagonist beta-funaltrexamine (beta-FNA) to investigate their ability to block the acute opioid-like effects of a single electroconvulsive shock (ECS). lCl, but not beta-FNA, attenuated the increase in hot-plate escape latencies seen following ECS. From these data, we conclude that ECS activates endogenous opioids which act upon delta rather than mu receptors to result in an increase in hot-plate escape latencies.