Impaired immune responses in streptozotocin-induced type I diabetes in mice. Involvement of high glucose.

Diabetes is widely believed to predispose to serious infections. However, the mechanisms linking diabetes and immunosuppression are not well defined. One potential mediator of the altered defence mechanisms is hyperglycaemia. It has been identified as the main factor contributing to the development of diseases associated with diabetes mellitus. In this study we analyse the immune response in diabetes and the direct effect of hyperglycaemia on T and B lymphocyte reactivity. Diabetes induced an early decrease in IgG levels in the secondary response. However, both primary responses against a T-cell-dependent or independent antigen were affected after 6 months of diabetes induction. T- and B- cell proliferation was only decreased at this time. To gain insight into the potential mechanisms involved, we evaluated the influence of hyperglycaemia over the immune response. Pre-incubation of lymph node and spleen cells in a high glucose (HG) containing medium led to a significant time- and dose-dependent decrease in T- and B-cell proliferation. This effect was associated with the presence of HG-derived supernatants. Still viable cells after HG exposition were able to improve their proliferative response when cultured with the mitogen in a fresh standard medium. HG diminished cell viability, increased apoptosis and induced oxidative stress in lymphocytes. These results indicate that HG concentrations can directly affect lymphoid cell growth. An increase in oxidative stress would be implicated in this deleterious effect. The possibility that prolonged exposure to pathologically HG concentrations would result in the immunosuppressive state observed in diabetes is also discussed.

A novel BCR-ABL rearrangement in a Philadelphia chromosome-positive chronic myelogenous leukaemia variant with thrombocythaemia.

Reverse transcription polymerase chain reaction was used to detect an unusual BCR/ABL transcript in a patient who presented with thrombocythaemia and was Philadelphia chromosome positive but weakly reactive to conventional BCR/ABL fusion probes. Sequencing revealed the presence of a 12 bp insert between BCR exon2 (b2) and ABL exon2 (a2). In such cases the use of conventional BCR/ABL probes may lead to false negative results. This is the first report of this transcript. Clinically the patient has responded well to therapy.

Bone marrow transplantation depleted of T cells followed by repletion with incremental doses of donor lymphocytes for relapsing patients with chronic myeloid leukemia: a therapeutic strategy.

BACKGROUND: For patients with chronic myeloid leukemia (CML), long-term survival after stem cell transplantation requires adequate control of graft-versus-host disease (GVHD) and disease recurrence. Relapsing patients respond to donor lymphocyte infusion (DLI) but develop life-threatening complications. METHODS: Patients with CML in first chronic phase received bone marrow (n = 14) or peripheral blood progenitor cell transplants (n = 4) from HLA-identical siblings. GVHD prophylaxis was by ex vivo T-cell depletion with CAMPATH 1G. If disease recurred, donors' mononuclear cells were collected by apheresis, the CD3 samples commencing at 10(6)/kg were aliquoted at half-log increment intervals, cryopreserved, and infused until disease clearance. RESULTS: Eighteen patients (median age: 32.5 years) received transplants. All engrafted without procedure-related mortality. Fourteen patients relapsed, and 13 entered the DLI program. Two developed extensive GVHD after single schedule infusions ranging from 89x10(6) to 670x10(6) mononuclear cells/kg, and one survives in complete remission (CR). The rest, treated with incremental dose DLI, experienced no acute toxicities. One, who had developed grade III steroid-responsive GVHD, died in CR2 from opportunistic infections. Steroids reversed limited cutaneous GVHD and elevated liver enzymes in five patients. Three others developed pancytopenia, and two restored blood counts only after donor peripheral blood progenitor cell infusions. Molecular CR2 was established in 12/13 patients, occurring in 10/11 (91%) on the incremental program at a median accumulation of 67 (range: 5-166) x10(6) CD3 cells/kg. Sixteen of 18 (89%) survive at median of 854.5 days from bone marrow transplantation, 4 in CR1 and 10 in CR2 at a median disease-free survival (for remission 2) duration of 341 days. The median combined disease-free survival of the 14 patients in CR 1+2 is 660 days, with 99% average performance status. CONCLUSIONS: Escalating DLI leads to safe new molecular CR in most CML relapse patients. These results raise the possibility of using "safe" transplantation programs of T-cell depletion, that include graded DLI as prevention against disease recurrence.

A procedure for RT-PCR amplification of mRNAs on histological specimens.

Detection in combination with localization of low copy gene expression can be difficult to achieve. The use of reverse transcription PCR on tissue sections with a fluorescent marker provides localization of mRNA expression on a cellular level, and when combined with confocal microscopy and image analysis it also allows for an estimate of the relative intensity of fluorescence. A good example of the application of this method is the localization of nerve growth factor (NGF) mRNA expression in the inner ear. NGF through indirect tests appears to be present in this system, yet NGF mRNA could not be localized with in situ hybridization using radiolabeled riboprobes. Using fluorescent in situ RT-PCR, we can easily detect the presence of NGF mRNA and localize it to specific cell types within the maturing inner ear.

Videodensitometric quantitation of aortic regurgitation by digital subtraction aortography using a computer-based method analyzing time-density curves.

To assess the clinical role of computer analysis of time-density curves in the evaluation of aortic regurgitation (AR), digital subtraction aortography (DSA) and cineaortography were performed sequentially in 17 patients with varying degrees of AR (1+ to 4+) and in 4 control patients. DSA was performed at a rate of 30 frames/s on a 512 X 512 X 8 bit pixel matrix using the same total volume and injection rate, but with half the amount of contrast agent as standard cineaortography. A 30 X 30 pixel area of interest was identified in the aorta above the valve plane and in the left ventricle where the AR stream was seen. The density of both areas of interest and the ratio of left ventricular/aortic area of interest density was calculated in each frame and then plotted vs time. The ratio at the end of injection (LVd/Aod) had an excellent correlation with cineaortography (chi 2 = 19, p less than 0.001), ranging from 0 to 0.2 in patients with no AR, 0.2 to 0.5 in those with 1+ AR, 0.5 to 0.7 in those with 2+ AR, 0.7 to 0.9 in those with 3+ AR and more than 0.9 in those with 4+ AR. Thus, quantitative assessment of AR by computer analysis of time-density curves derived from DSA is a new and objective technique with significant clinical potential.

Inhibition of choline efflux results in enhanced acetylcholine synthesis and release in the guinea-pig corticocerebral synaptosomes.

Synthesis and release of [3H]acetylcholine ([3H]ACh) were measured in synaptosomes from the guinea pig cerebral cortex after preloading with [3H]choline ([3H]Ch). We demonstrate here that inhibition of choline (Ch) efflux results in an increase in acetylcholine (ACh) synthesis and release. Our findings are as follows: (1) inhibition of [3H]Ch efflux by hemicholinium-3 (HC-3) (100 microM), increased the levels of both the released (116% of control) and the residing (115% of control) [3H]ACh. (2) The muscarinic agonist, McN-A-343 (100 microM), which was previously shown to inhibit Ch efflux, also increased the released (121% of control) and the residing (109% of control) [3H]ACh. (3) Omission of Na+ ions (which are required for Ch transport) from the incubation medium had similar effects to those observed with McN-A-343 and HC-3. These results suggest inverse relationships between Ch efflux on one hand, and ACh synthesis and release on the other hand. (4) Depolarization with 50 mM K+, or with the K+ channel blocker, 4-aminopyridine (100 microM), also increased the total level of [3H]ACh (113 and 107% of nondepolarized synaptosomes, respectively). However, whereas conditions that inhibit Ch transport such as HC-3, McN-A-343 and "no sodium" increased both the residing and the released [3H]ACh depolarization with high K+ or 4-aminopyridine reduced the residing (79 and 87% of control, respectively) and increased only the released [3H]ACh (182 and 148% of control, respectively).(ABSTRACT TRUNCATED AT 250 WORDS)

Towards a public health approach to bioethics.

In this paper we examine the central commitments of bioethical enquiry and reasoning from a public health perspective. We argue that a core element of American national culture is individualism, which resonates in scholarly and popular debates. Our contention is that the habitus of bioethical debate is in large measure animated by an overriding concern with the individual, and the resulting social practice of the community has been to downplay the importance and legitimacy of group-level health care dilemmas. This paper calls for re-focusing of bioethics by employing a public health perspective, which would include a population focus, evidence-based research topics, and engagement of the ethical dilemmas that arise from decisions concerning prevention. Racial and ethnic health disparities throughout the life span of a population in central New York State are used to illustrate the need for a public health focus in bioethics.

Histamine-mediated acetylcholine release in the guinea-pig ileum.

The isometric contraction induced by histamine in guinea-pig ileum longitudinal muscle was biphasic in Krebs ([Ca]: 3.36 mM) but monophasic in Tyrode solution ([Ca]: 1.80 mM). The late phase (histamine: 20-400 nM) was common to the two solutions but the early phase (histamine: 2-20 nM) was observed only in Krebs solution. This early phase could be blocked with atropine (0.01-0.2 microM), morphine (0.1, 1 microM), adenosine (5, 20 microM) and tetrodotoxin (0.3 microM) without affecting the late phase. Washout of morphine or adenosine was fast. Neostigmine (100 nM) greatly potentiated the effect of histamine (4 nM) in the early phase, the muscle undergoing almost maximum contraction but also reversible desensitization to doses of histamine less than or equal to 20 nM for as long as 40 min after washout. Beyond this concentration, the preparation responded to increasing doses of histamine as observed in the late phase. It is concluded that low concentrations of histamine that have no observable direct effect on muscle contractility release acetylcholine in the presence of [Ca] 3.36 mM, the early phase being entirely due to release of endogenous acetylcholine.

Affinity of nortriptyline to muscarinic receptors in the bladder and ileum of man and guinea-pig.

The antagonism by nortriptyline of carbachol- or urecholine-induced contractions was studied in strips of ileum and bladder derived from man and guinea-pig. Analyses of the results by the dose ratio method (Schild plots) showed significant differences in the affinities of the relevant muscarinic receptors to the antagonist: The Ki values in microM were as follows: Human ileum, 0.938; human bladder, 0.298; guinea-pig ileum, 0.159; guinea-pig bladder, 0.333 and 0.453. In man, the higher affinity of the drug to the receptors in the bladder than to those in the ileum may be of consequence in its therapeutic application as an antienuretic agent.

Acetylcholine mediation of the contractile response to histamine in human bladder detrusor muscle.

In Krebs solution, histamine evokes in human bladder detrusor muscle strips a dose-dependent contractile response which consists of two pharmacologically distinct responses: a high-sensitivity response evoked at 0.4-2 microM histamine, which is potentiated by neostigmine (0.1 microM) or blocked by atropine (0.1 microM) or ranitidine (1 microM); a low-sensitivity response evoked at 4-40 microM histamine and blocked by dimethindene or diphenhydramine. These findings suggest that the contractile response to low doses of histamine is mediated by acetylcholine released from a site proximal to the muscle. This effect of histamine seems to be mediated by a site which is insensitive to the H1 antagonists dimethindene and diphenhydramine but blocked by the H2 antagonist ranitidine.

Antihistaminic properties of AF-14, an experimental quinuclidine derivative: discrimination between two histaminergic sites in both guinea-pig ileum and bladder.

AF-14 (1-aza-4-phenyltricyclo[6.2.2.0(2,7)]dodecan-5-one), a selective antimuscarinic agent, was shown to block the histamine-induced contractile response in Krebs solution in guinea-pig ileum and bladder. Careful linear regression analyses of the concentration-response relationship revealed that AF-14 antagonism consisted of two sequential and distinct phases, corresponding to an early phase and a late phase of the contractile response to histamine. In the late phase, the action of AF-14 was consistent with competitive antagonism, its pA2 (95% confidence limits) being 5.80-6.06 (ileum) and 5.66-5.80 (bladder). In the early phase, AF-14 almost completely blocked the contractile response at a concentration of 100-300 nM, which was much less than required to block the late phase or cholinergic contractions of the ileum or bladder. It is concluded that AF-14 discriminates between two histamine-sensitive sites that mediate muscle contraction in each of the two organs.

A study of the contractile response to acetylcholine in human ileal and detrusor muscle: origin of the low efficacy of acetylcholine.

In Krebs solution (3.36 mM Ca2+), the maximal contractile response of human ileal and urinary bladder detrusor muscle to acetylcholine (ACh) was 40-60% that to carbachol (CCh). The maximum response to ACh was reached at a bath concentration of about 1 microM and was maintained throughout a range extending to 100 microM. In the presence of neostigmine (0.1 microM), the maximum response to ACh reached the level of that of CCh. However, bioassay of bath concentrations of ACh at various points of the maximal response in the absence of neostigmine revealed only slight to insignificant diminution of the applied concentration of ACh. Joint application of ACh and CCh generated a dose-response profile consistent with a model of competitive antagonism between a full agonist (CCh) and a partial one (ACh). Also, choline (100 microM) reduced the maximum response to ACh in the presence of neostigmine and that to CCh to 60-80% of control. These observations are consistent with a mechanism whereby intact cholinesterase together with its substrate ACh and possibly a breakdown product of ACh constitute a filter or diffusional barrier regulating the flow of agonist from the enzyme compartment to the receptor compartment.

Older men's health. Sociocultural and ecological perspectives.

This article identifies a set of conditions that renders the morbidities and earlier deaths of men as the outcome. The article also discusses four factors that affect older men's health and longevity: culture, class, race and ethnicity, and social organization and participation.

Utility of relative cerebral blood volume mapping derived from perfusion magnetic resonance imaging in the routine follow up of brain tumors.

It was recently demonstrated that imaging of brain tumors by relative cerebral blood volume (CBV) maps reconstructed from dynamic magnetic resonance (MR) data provide similar diagnostic information compared to positron emission tomography (PET) or 201Tl single-photon emission computerized tomography (201Tl-SPECT) scans. The authors used relative CBV mapping for routine follow-up evaluation of patients with brain tumors and compared its sensitivity to diagnostic MR imaging, 201Tl-SPECT and clinical assessment. Fifty-nine patients were prospectively followed using 191 concomitant studies of dual section relative CBV maps, MR imaging, 201Tl-SPECT, and neurological evaluations. Studies were repeated every 2 to 3 months (median three evaluations/patient). The relative CBV maps were graded as relative CBV 0 to 4, where Grades 3 and 4 are indicative of proliferating tumors (four = rapid leak). There were 44 high-grade and 15 low-grade tumors followed during treatment. During the follow-up period a change in relative CBV grade was observed in 56% of the patients, revealing an increasing grade in 72% of them. The rapid leak phenomenon was detected in 35% of all studies and in 81% of those with a worsening relative CBV grade. Tumor progression was detected earlier by relative CBV maps as follows: earlier than MR imaging in 32% of the studies (earlier by a median of 4.5 months; p < 0.01); earlier than 201Tl-SPECT in 63% (median 4.5 months; p < 0.01), and earlier than clinical assessment in 55% (median 6 months; p < 0.01). In 82% of studies with positive MR imaging but negative 201Tl-SPECT, the lesions were smaller than 1.5 cm. The relative CBV maps clearly delineated the appearance of rapid leak in these lesions. Routine use of relative CBV maps that can be implemented on any high-field MR unit and added to the regular MR evaluation provides useful functional information in patients with brain tumors. When used as an adjunct follow-up evaluation it proved more sensitive than the other modalities for early prediction of tumor growth. It is very sensitive to small regional changes, unlike functional imaging such as PET or SPECT scans. Based on previous experience with 76 regional CBV studies, the authors conclude that regional CBV mapping correlates with active tumor and it may separate enhancing scar and radiation injury from infiltrative tumor. A new effect named the rapid leak phenomenon was also observed; this phenomenon, as identified on the regional CBV maps, correlates with high malignancy.

In vivo assessment of the window of barrier opening after osmotic blood-brain barrier disruption in humans.

OBJECT: Osmotic blood-brain barrier (BBB) disruption induced by intraarterial infusion of mannitol is used in conjunction with chemotherapy to treat human brain tumors. The time course to barrier closure, or the so-called therapeutic window, has been examined in animals but little information is available in humans. The authors, therefore assessed the time course to barrier closure after osmotic BBB disruption in humans. METHODS: Disruption of the BBB was demonstrated using 99mTc-glucoheptonate (TcGH) single-photon emission computerized tomography (SPECT) scanning in 12 patients who were treated monthly with combination chemotherapy in conjunction with BBB disruption. The primary diagnosis was primary central nervous system lymphoma in seven patients and primitive neuroectodermal tumors in five. The TcGH (20 mCi) was injected at 1- to 480-minute intervals after osmotic BBB disruption, and patients underwent SPECT scanning after 4 hours. A total of 38 studies was performed. Good-to-excellent BBB disruption was obtained in 29 procedures and poor-to-moderate disruption was seen in the other nine studies. The TcGH indices correlated with the degree of BBB disruption as measured postprocedure on contrast-enhanced CT scans (r = 0.852). Mean baseline TcGH indices were 1.02+/-0.07. For the group of patients with good-to-excellent disruptions the mean indices at 1 minute postdisruption measured 2.19+/-0.18. After 40 minutes no significant change was noted (mean index 2.13+/-0.2). Then the indices declined more steeply and at 120 minutes after the disruption the index was 1.36+/-0.02. A very slow decline was noted between 120 and 240 minutes after mannitol infusion. At 240 minutes the barrier was still open for all good-to-excellent disruptions (index 1.33+/-0.08) but at 480 minutes the mean indices had returned to the baseline level. CONCLUSIONS: Results of these in vivo human studies indicate that the time course to closure of the disrupted BBB for low-molecular-weight complexes is longer than previously estimated. The barrier is widely open during the first 40 minutes after osmotic BBB disruption and returns to baseline levels only after 6 to 8 hours following the induction of good or excellent disruption. These findings have important clinical implications for the design of therapeutic protocols.

A comparative study of the affinities of some tricyclic antidepressants for the muscarinic cholinergic receptor in human and guinea-pig bladder, ileum and brain in relation to differential drug potency.

Following a report that nortriptyline was found useful in the control of enuresis in adults, presumably as an anticholinergic, its likely mechanism of action and apparent bladder specificity have now been investigated in vitro. The ratios of anticholinergic potencies (reciprocal of dissociation contents, Ki) for four different tricyclic antidepressants, derived from competitive binding assays with (-)[3H]QNB in tissue homogenates, in the order (human) detrusor muscle/ileal longitudinal muscle/caudate, are as follows: Nortriptyline, 5/4/7; desipramine, 2/1/5/; clomipramine, 4/3/27; amitriptyline, 25/14/56. The apparent selective effect of nortriptyline on the bladder cannot be ascribed to its higher affinity to bladder receptors. Still, this drug is the least discriminatory of the four. Hence, at a given concentration, it is expected to affect tissue embodying a low density receptor pool sooner than tissue having a large receptor reserve. The ratios of the densities of (-)[3H]QNB binding sites in the order detrusor muscle/ileal muscle/cortex is 1/3/5, supporting the present contention. In the guinea-pig, the ratios of the anticholinergic potency in the order bladder/proximal ileum/distal ileum/cortex are as follows: Nortriptyline, 25/5/6/33; desipramine, 8/2/2/14; amitriptyline, 100/14/20/100; clomipramine, 17/3/5/33. Also, the ratios of the densities of binding sites are 1/6/5/2. Hence, data derived from assays in the guinea-pig are not representative of those derived from human tissue.

"Breaking the bureaucracy": drug registration and neocolonial relations in Egypt.

According to the Egyptian Ministry of Health, the per capita use of prescription drugs in Egypt is amongst the highest in the world. Multinational pharmaceutical companies license their proprietary products for manufacture and sale in Egypt through their Egyptian subsidiaries. A Ministry of Health Committee reviews and approves for sale all drugs marketed in the country. Aside from being an extremely lucrative market itself, approval of a drug for sale and manufacture in Egypt also opens to the pharmaceutical companies other markets in the Arab world. The Egyptian drug approval process is thus both important for assuring the health of Egyptian nationals and a high-stakes activity for the pharmaceutical companies. This paper examines the social relations and interactions of multinational pharmaceutical representatives in Egypt with Egyptian researchers in relation to the Ministry of Health's drug approval process. From time-to-time events focus attention on the huge financial rewards reaped by multinational pharmaceutical companies from their activities in lesser developed countries. This attention not infrequently has revealed the "drugging of the Third World" as a result of actions by expatriate multinational pharmaceutical officials. Indigenous review procedures such as those established by the Egyptian Ministry of Health might guard against such external exploitation. This paper shows how in place of external exploitation, indigenous pharmaceutical company officials have manipulated local patterns of social interaction to construct a system of reciprocal obligations which may frustrate intended safeguards, and by reconstructing colonial institutional structures, creates a pattern of neocolonialism in Egypt.

Frey syndrome: treatment with temporoparietal fascia flap interposition.

There is a 10% to 48% reported incidence of clinically significant gustatory sweating after parotid surgery or injury. Various medical and surgical treatments have been used in the attempt to treat this socially embarrassing condition. These treatments are not always effective and often have unwanted risks and adverse effects. They also do not address the post-parotidectomy defect. Prevention of Frey syndrome and correction of the postoperative contour deformity after parotidectomy have recently been achieved by interposition of temporoparietal fascia flap between the parotid gland and the cheek skin flap at the time of parotidectomy. This article presents the first report (to our knowledge) of an established case of Frey syndrome being treated with temporoparietal fascia flap interposition.

A path not taken: a cultural analysis of regrets and childlessness in the lives of older women.

This paper explores regrets about childlessness in 90 older women interviewed using qualitative methods. Regrets were discussed in the context of the changing meaning of childlessness over the life course. We found that issues of regret are situated in a cultural system that renders childless women marginal. We argue that regrets should be understood in a wider cultural context that incorporates the cultural construction of the self over time.