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Biomedical subjects

N Syed

Publications and source records attributed to N Syed.

At least 19 recordsLinked to original sources

Evaluation of retinal photoreceptors and pigment epithelium in a female carrier of choroideremia.

PURPOSE: To clarify the pathogenesis of choroideremia. STUDY DESIGN: Human tissue study. TISSUES: Eyes of an 88-year-old symptomatic female carrier of choroideremia (CHM) and six normal, age-matched donors. METHODS: The eyes were processed for histopathologic examination, including immunocytochemistry with an antibody against the CHM gene product, REP-1, and retinal cell-specific markers. RESULTS: The CHM carrier retina showed patchy degeneration, but the photoreceptor and retinal pigment epithelium (RPE) loss appeared to be independent. The choriocapillaris was normal except where retinal areas were severely degenerate. The CHM gene product, REP-1, was localized to the cytoplasm of rods but not cones. CONCLUSIONS: It has generally been considered that photoreceptor degeneration in CHM is secondary to loss of the choriocapillaris or RPE. This study suggests that the rod photoreceptors are a primary site of disease in CHM.

Adaptor Proteins, Signal Transducing↗

Eradication of house dust mite from homes of atopic asthmatic subjects: a double-blind trial.

BACKGROUND: House dust mite (HDM) allergens can accumulate to very high levels in homes. From the observed sensitivity of HDMs to heat and their allergens to steam, a novel treatment of furnishings has been developed. OBJECTIVE: We sought to determine whether combined steam and heat treatment of home furnishings reduced asthmatic patients' bronchial hyperreactivity (BHR) and lowered HDM antigen loads. METHODS: The homes of 30 asthmatic subjects aged 18 to 45 years were randomly allocated into 3 groups. In groups 1 and 2 mattresses and duvets were treated with hot air (110 degrees C), followed by steam and then heat again. All their carpets were steam cleaned. Group 2 also had a special ventilation system installed above each patient's bedroom. The homes of subjects in group 3 were sham treated. Neither patient nor laboratory staff was aware of the types of treatment. Der p 1 and 2 levels in the household dust from the lounge, bedroom carpet, and beds were determined before and after treatment and then at 6 and 12 months. BHR, measured by using histamine PD(20) values, was recorded during the 4-week run-in period and at 3, 6, 9, 12 months after treatment. RESULTS: Active heat-steam treatment of homes caused a sustained reduction of Der p 1 (P =.003) and Der p 2 (P =.001) compared with no change in sham-treated group 3 homes. Patients whose homes were treated showed a 4-fold reduction in BHR at 9 months in group 1 and throughout the posttreatment period in group 2. No change was observed in the asthmatic subjects whose homes were not treated. These improvements were sustained for 12 months in the homes with bedroom ventilation units. CONCLUSIONS: A single treatment of home furnishings reduced mite allergen load to below the risk level for sensitization and improved the asthmatic patients' BHR by 4-fold.

Adolescent↗

A fast and efficient method for transiently transfecting ES cells: application to the development of systems for conditional gene expression.

Classically, mouse embryonic stem (ES) cells are transfected by electroporation, a method that requires a large number of cells. Here we describe a protocol using a liposome based transfection agent that is a very simple, rapid and cost effective way of transiently transfecting very low numbers of ES cells. We found this method very useful in screening a large number of ES clones when working with inducible expression systems in which at least two elements are required for regulated expression of the gene of interest. After stable transfection of the first component, clones can be easily and rapidly screened for expression of the gene of interest by transiently transfecting the second component of the system using this protocol.

Cells, Cultured↗

Learning, memory and a respiratory central pattern generator.

In an attempt to elucidate the causal mechanisms underlying learning and memory we have developed a model system, aerial respiration in the pond snail Lymnaea stagnalis. A three-neuron central pattern generator (CPG) whose sufficiency and necessity have been demonstrated mediates this behaviour. Aerial respiration, while an important homeostatic behaviour, is inhibited by the activation of the whole body withdrawal response that the animal uses to protect itself. We found that it was possible to operantly condition snails not to perform aerial respiration in a situation, a hypoxic environment, where aerial respiration should predominate. Operant conditioning was achieved by eliciting the pneumostome withdrawal response, part of the whole body withdrawal response, each time the animal attempted to open its pneumostome to breathe. Yoked control animals did not demonstrate an alteration in breathing behaviour. Subsequently we determined neural correlates of this associative behaviour and found that neuronal changes are distributed throughout the CPG. This preparation may afford us the opportunity to determine the casual neuronal changes that underlie learning and memory of associative conditioning.

Animals↗

Ten years' experience with enteral levodopa infusions for motor fluctuations in Parkinson's disease.

We report our long-term experience using enteral levodopa infusions in 22 patients with Parkinson's disease and severe motor fluctuations. Amelioration of intractable dyskinesias was the most important factor that determined whether patients chose to continue using the infusion pump system. Mechanical and physical problems associated with enteral access were the most common reasons for which patients discontinued pump use. Nearly all patients continued to have dramatically increased on time for the duration of follow up, suggesting that technically less-cumbersome systems that provide continuous dopaminergic stimulation are worthwhile and should be developed.

Administration, Oral↗

Renal actions of endothelin-1 under endothelin receptor blockade by BE-18257B.

PURPOSE: Endothelin-1 (ET-1), a peptide produced by the vascular endothelium, causes profound renal vasoconstriction by binding to ET-A receptors. The present study examined the renal actions of ET-1 after ET-A receptors were blocked by BE-18257B to unmask the functions of ET-B receptors. MATERIALS AND METHODS: Renal hemodynamics and clearance measurements were obtained in anesthetized dogs after intrarenal infusion of BE-18257B at 100 ng./kg./min. (Group 1), after intrarenal infusion of ET-1 at 2 ng./kg./min. (Group 2), or after intrarenal infusion of ET-1 superimposed on BE-18257B (Group 3). RESULTS: In Group 1, BE-18257B infusion did not alter arterial pressure, renal blood flow (RBF), GFR or tubular function. In Group 2, ET-1 infusion led to a significant decrease in RBF and GFR (37 and 40%, respectively) without altering arterial pressure. Urinary volume and sodium excretion were not changed but osmolality decreased significantly. In Group 3, BE-18257B infusion significantly attenuated the decrease in RBF caused by ET-1 and increased GFR by 40% without altering arterial pressure, associated with significant diuresis and natriuresis. CONCLUSION: Renal vasoconstriction caused by ET-1 is attenuated by ET-A receptor blockade with BE-18257B, which unmasks the hemodynamic and tubular actions of ET-B receptors. As a result, it limits the ET-1 induced decrease in RBF and raises GFR, and leads to a diuresis and natriuresis.

Anesthesia↗

Retinoblastoma in an eye with congenital uveal coloboma.

Retinoblastoma and congenital ocular abnormalities rarely coexist. The appearance of an intraocular tumor in a child with preexisting leukokoria may make the diagnosis of retinoblastoma more challenging. This report describes a unique case of unilateral congenital uveal coloboma and ipsilateral retinoblastoma in a patient with no family history of either abnormality and normal cytogenetic analysis.

Blotting, Southern↗

Ciliary neurotrophic factor, unlike nerve growth factor, supports neurite outgrowth but not synapse formation by adult Lymnaea neurons.

The nerve growth factor (NGF) family and ciliary neurotrophic factor (CNTF) support survival and/or neurite outgrowth of many cell types. However, it is not known whether the neurite outgrowth induced by neurotrophic factors results in the formation of synapses. We tested NGF and CNTF for their ability to induce neurite outgrowth and synapse formation in vitro by interneurons from the mollusc Lymnaea. Dopaminergic and peptidergic interneurons survived in the absence of neurotrophic factors but exhibited robust outgrowth in response to both NGF and CNTF. Chemical synapses formed between these interneurons and their target neurons cultured in NGF, but synapses were absent in CNTF. Survival, neurite outgrowth, and synaptogenesis are therefore differentially regulated in these neurons.

Age Factors↗

Rhythmic activities of isolated and clustered pacemaker neurons and photoreceptors of Aplysia retina in culture.

Each eye of Aplysia contains a circadian clock that produces a robust rhythm of optic nerve impulse activity. To isolate the pacemaker neurons and photoreceptors of the eye and determine their participation in the circadian clock and its generation of rhythmic autoactivity, the retina was dissociated and its cells were placed in primary cell culture. The isolated neurons and photoreceptors survived and vigorously extended neurites tipped with growth cones. Many of the photoreceptors previously described from histological sections of the intact retina were identified in culture, including the large R-type photoreceptor, which gave robust photoresponses, and the smaller tufted, whorled, and flared photoreceptors. The pacemaker neurons responsible for the rhythmic impulse activity generated by the eye were identified by their distinctive monopolar morphology and recordings were made of their activity. Isolated pacemaker neurons produced spontaneous action potentials in darkness, and pacemaker neurons attached to fragments of retina or in an isolated cluster interacted to produce robust spontaneous activity. This study establishes that isolated retinal pacemaker neurons retain their innate autoactivity and ability to produce action potentials in culture and that clusters of coupled pacemaker neurons are capable of generating robust autoactivity comparable to pacemaker neuron rhythmic activity recorded in the intact retina, which was previously shown to correspond to 1:1 with the optic nerve compound action potential activity.

Action Potentials↗

Operant conditioning of aerial respiratory behaviour in Lymnaea stagnalis

In this study, we operantly conditioned the aerial respiratory behaviour of the freshwater snail Lymnaea stagnalis. Aerial respiration in Lymnaea stagnalis is accomplished by the spontaneous opening and closing of its respiratory orifice, the pneumostome, at the water surface. Weak tactile stimulation of the pneumostome area, when the pneumostome is open, evoked only the pneumostome closure response, which is one aspect of the escape-withdrawal reflex. Pneumostome stimulation resulted in its closure and the termination of aerial respiratory activity. A contingent tactile stimulation paradigm was used to operantly condition the animals. Stimulation of the pneumostome whenever the animal attempted to breathe resulted in significantly fewer attempts to open the pneumostome as training progressed. The latency of the first breath (subsequent to stimulation), the number of breaths and the total breathing time were measured before and after each training period. Significant, quantifiable changes in these behavioural parameters were observed only in the operant conditioning group animals. Control animals receiving tactile stimulation to their pneumostome not contingent upon pneumostome opening movements (yoked controls) or those that were physically prevented from surfacing to breathe (hypoxic controls), did not exhibit significant changes in these behavioural parameters. Our data provide the first direct evidence for operant conditioning of respiration in any animal.

Journal Article↗

Inhibition of the respiratory pattern-generating neurons by an identified whole-body withdrawal interneuron of Lymnaea stagnalis

Respiration and the whole-body withdrawal are two incompatible behaviors in the freshwater snail Lymnaea stagnalis. Whole-body withdrawal behavior is believed to be higher on the behavioral hierarchy than respiratory behavior. A central pattern generator (CPG) underlies respiratory behavior; whole-body withdrawal is mediated by a network of electrically coupled neurons. In this study, we provide evidence that the behavioral hierarchy between the whole-body withdrawal and the respiratory behaviors is established at the interneuronal level. We demonstrate that an identified whole-body withdrawal interneuron inhibits both muscular and neuronal components of the respiratory behavior in Lymnaea stagnalis. A pair of identified, electrically coupled interneurons, termed left and right pedal dorsal 11 (L/RPeD11), coordinates the whole-body withdrawal behavior in Lymnaea stagnalis. In the present study, RPeD11 inhibited spontaneously occurring respiratory CPG activity in isolated brain preparations. In addition, electrical stimulation of RPeD11 in a semi-intact preparation also inhibited respiratory CPG interneuron RPeD1. The synaptic connections between RPeD11 and the respiratory CPG neurons RPeD1 and visceral dorsal 4 (VD4) persisted in the presence of high-Ca2+/high-Mg2+ saline, suggesting the possibility that they may be monosynaptic. In a semi-intact preparation (lung­mantle, pneumostome and central nervous system), electrical stimulation of RPeD11 induced pneumostome and columellar muscle contractions while inhibiting the activity of RPeD1. Moreover, mechanical stimulation of the respiratory orifice, the pneumostome, excited RPeD11, while its effects on the respiratory CPG neuron (RPeD1) were inhibitory. To determine the monosynaptic nature of connections between RPeD11 and the respiratory CPG neurons in the intact nervous system, we constructed these synapses in culture. RPeD11 and individual respiratory interneurons were isolated from their respective ganglia and co-cultured under conditions that support neurite outgrowth. Following neuritic overlap, RPeD11 was found to establish inhibitory synapses with the respiratory interneurons, supporting the hypothesis that these synaptic connections are likely to be monosynaptic in the intact central nervous system.

Journal Article↗

DIFFERENTIAL TRACER COUPLING BETWEEN PAIRS OF IDENTIFIED NEURONES OF THE MOLLUSC LYMNAEA STAGNALIS

Electrical coupling is a common means of cell-to-cell communication in both neuronal and non-neuronal tissues (Lowenstein, 1985). Within the nervous system, many electrically coupled neurones exhibit dye coupling (Bennett, 1973; Stewart, 1978; Glantz and Kirk, 1980; Spencer and Satterlie, 1980; Fraser and Heitler, 1993); however, some electrically coupled cells do not dye-couple (Audesirk et al. 1982; Murphy et al. 1983; Berdan, 1987; Robinson et al. 1993; Veenstra et al. 1993). Electrical coupling and dye coupling, often considered in parallel, are in fact two different parameters that can vary independently (e.g. Audesirk et al. 1982; Perez-Armendariz et al. 1991). The giant identified neurones of pulmonate and opisthobranch molluscs have frequently been used for studies of neuronal communication and its plasticity (Winlow and McCrohan, 1987; Bulloch, 1989). In the present study, we explored the relationship between electrical and tracer coupling in both strongly and weakly coupled pairs of molluscan neurones. Specifically, we examined electrically coupled, identified neurones in a freshwater pond snail, Lymnaea stagnalis L., and tested for tracer coupling with Lucifer Yellow CH and biocytin. The cells examined were the strongly electrically coupled neurones, visceral dorsal 1 (VD1) and right parietal dorsal 2 (RPD2) (Boer et al. 1979; Benjamin and Pilkington, 1986), and the weakly coupled neurones, left buccal 1 (LB1) and right buccal 1 (RB1) (Benjamin and Rose, 1979). The use of these particular neurones made it possible to compare electrical coupling with tracer coupling in the molluscan central nervous system (CNS). All experiments were performed on laboratory-bred Lymnaea stagnalis (Mollusca, Pulmonata), maintained as previously described (Ridgway et al. 1991). The CNS was dissected from mature animals (16­18 mm shell length) and pinned to the silicone rubber (RTV 616 GE) base of a recording dish in normal saline (51.3 mmol l-1 NaCl, 1.7 mmol l-1 KCl, 4.1 mmol l-1 CaCl2, 1.5 mmol l-1 MgCl2 and 5 mmol l-1 Hepes, pH 7.9). Following removal of the outer connective tissue sheath, a small Pronase crystal (Sigma, type XIV, P-5147), held by forceps, was carefully applied to specific ganglia; this treatment softened the inner sheath and facilitated microelectrode penetration. The CNS was then rinsed several times at 5 °C in normal saline.

Journal Article↗

Followup prostate biopsy in patients with carcinoma of the prostate treated by 192iridium template irradiation plus supplemental external beam radiation.

We reviewed our experience with 81 patients who had undergone followup needle biopsies of the prostate between 12 and 27 months after 192iridium template radiotherapy combined with external beam radiation therapy. When broken down by stage 82% of the patients with stage A2, 92% with B1, 95% with B2 and 55% with C disease demonstrated a negative biopsy and adequate local control of the cancer. We believe that in elderly patients, poor risk patients with early stage disease and stage C lesions this low morbidity, low mortality therapy offers a viable option to other modalities of treatment.

Aged↗

Circulating Epstein-Barr virus-carrying B cells in acute malaria.

Epstein-Barr virus (EBV) infection and Plasmodium falciparum malaria are two known cofactors in the aetiology of endemic Burkitt's lymphoma. To assess the relation between these factors, limiting dilution analysis was used to assess the number of EBV-carrying B cells in the circulation of Gambian children during and after acute malaria. Numbers of virus-carrying cells were five times higher in acute malaria patients and in UK patients with infectious mononucleosis than in convalescent malaria patients and in healthy control adults from the UK. Spontaneous outgrowth in limiting dilution cultures from acute malaria samples was inhibited by acyclovir, a viral DNA polymerase inhibitor. The mechanism of outgrowth, therefore, was virus release from the in-vivo infected cell, which led to infection and immortalisation of co-cultured normal B cells. The findings provide evidence that acute malaria is associated with an increase in the number of EBV-carrying B cells in the circulation. Because of this increase, there is a greater chance of a cytogenetic abnormality occurring in such a cell, with consequent evolution of Burkitt's lymphoma.

Acute Disease↗