Organ curvature sensing using pneumatically attachable flexible rails in robotic-assisted laparoscopic surgery.

In robotic-assisted partial nephrectomy, surgeons remove a part of a kidney often due to the presence of a mass. A drop-in ultrasound probe paired to a surgical robot is deployed to execute multiple swipes over the kidney surface to localise the mass and define the margins of resection. This sub-task is challenging and must be performed by a highly-skilled surgeon. Automating this sub-task may reduce cognitive load for the surgeon and improve patient outcomes. The eventual goal of this work is to autonomously move the ultrasound probe on the surface of the kidney taking advantage of the use of the Pneumatically Attachable Flexible (PAF) rail system, a soft robotic device used for organ scanning and repositioning. First, we integrate a shape-sensing optical fibre into the PAF rail system to evaluate the curvature of target organs in robotic-assisted laparoscopic surgery. Then, we investigate the impact of the PAF rail's material stiffness on the curvature sensing accuracy, considering that soft targets are present in the surgical field. We found overall curvature sensing accuracy to be between 1.44% and 7.27% over the range of curvatures present in adult kidneys. Finally, we use shape sensing to plan the trajectory of the da Vinci surgical robot paired with a drop-in ultrasound probe and autonomously generate an Ultrasound scan of a kidney phantom.

Palliative Lung Radiotherapy: Higher Dose Leads to Improved Survival?

AIMS: Choosing the optimal palliative lung radiotherapy regimen is challenging. Guidance from The Royal College of Radiologists recommends treatment stratification based on performance status, but evidence suggests that higher radiotherapy doses may be associated with survival benefits. The aim of this study was to investigate the effects of fractionation regimen and additional factors on the survival of palliative lung cancer radiotherapy patients. MATERIALS AND METHODS: A retrospective univariable (n = 925) and multivariable (n = 422) survival analysis of the prognostic significance of baseline patient characteristics and treatment prescription was carried out on patients with non-small cell and small cell lung cancer treated with palliative lung radiotherapy. The covariates investigated included: gender, age, performance status, histology, comorbidities, stage, tumour location, tumour side, smoking status, pack year history, primary radiotherapy technique and fractionation scheme. The overall mortality rate at 30 and 90 days of treatment was calculated. RESULTS: Univariable analysis revealed that performance status (P < 0.001), fractionation scheme (P < 0.001), comorbidities (P = 0.02), small cell histology (P = 0.02), 'lifelong never' smoking status (P = 0.01) and gender (P = 0.06) were associated with survival. Upon multivariable analysis, only better performance status (P = 0.01) and increased dose/fractionation regimens of up to 30 Gy/10 fractions (P < 0.001) were associated with increased survival. Eighty-five (9.2%) and 316 patients (34%) died within 30 and 90 days of treatment, respectively. CONCLUSION: In this retrospective single-centre analysis of palliative lung radiotherapy, increased total dose (up to and including 30 Gy/10 fractions) was associated with better survival regardless of performance status.

Dynamic physiological temperature and pressure sensing with phase-resolved low-coherence interferometry.

We report the development and characterisation of highly miniaturised fibre-optic sensors for simultaneous pressure and temperature measurement, and a compact interrogation system with a high sampling rate. The sensors, which have a maximum diameter of 250 µm, are based on multiple low-finesse optical cavities formed from polydimethylsiloxane (PDMS), positioned at the distal ends of optical fibres, and interrogated using phase-resolved low-coherence interferometry. At acquisition rates of 250 Hz, temperature and pressure changes of 0.0021 °C and 0.22 mmHg are detectable. An in vivo experiment demonstrated that the sensors had sufficient speed and sensitivity for monitoring dynamic physiological pressure waveforms. These sensors are ideally suited to various applications in minimally invasive surgery, where diminutive lateral dimensions, high sensitivity and low manufacturing complexities are particularly valuable.

The sympathetic innervation of the heart: Important new insights.

Autonomic control of the heart has a significant influence over development of life threatening arrhythmias that can lead to sudden cardiac death. Sympathetic activity is known to be upregulated during these conditions and hence the sympathetic nerves present a target for treatment. However, a better understanding of the anatomy and physiology of cardiac sympathetic nerves is required for the progression of clinical interventions. This review explores the organization of the cardiac sympathetic nerves, from the preganglionic origin to the postganglionic innervations, and provides an overview of literature surrounding anti-arrhythmic therapies including thoracic sympathectomy and dorsal spinal cord stimulation. Several features of the innervation are clear. The cardiac nerves differentially supply the nodal and myocardial tissue of the heart and are dependent on activity generated in spinal neurones in the upper thoracic cord which project to synapse with ganglion cells in the stellate complex on each side. Networks of spinal interneurones determine the pattern of activity. Groups of spinal neurones selectively target specific regions of the heart but whether they exhibit a functional selectivity has still to be elucidated. Electrical or ischemic signals can lead to remodeling of nerves in the heart or ganglia. Surgical and electrical methods are proving to be clinically beneficial in reducing atrial and ventricular arrhythmias, heart failure and severe cardiac pain. This is a rapidly developing area and we need more basic understanding of how these methods work to ensure safety and reduction of side effects.

Effect of electrical vs. chemical deep brain stimulation at midbrain sites on micturition in anaesthetized rats.

AIM: To understand how deep brain stimulation of the midbrain influences control of the urinary bladder. METHODS: In urethane-anaesthetized male rats, saline was infused continuously into the bladder to evoke cycles of filling and voiding. The effect of electrical (0.1-2.0 ms pulses, 5-180 Hz, 0.5-2.5 V) compared to chemical stimulation (microinjection of D,L-homocysteic acid, 50 nL 0.1 M solution) at the same midbrain sites was tested. RESULTS: Electrical stimulation of the periaqueductal grey matter and surrounding midbrain disrupted normal coordinated voiding activity in detrusor and sphincters muscles and suppressed urine output. The effect occurred within seconds was reversible and not secondary to cardiorespiratory changes. Bladder compliance remained unchanged. Chemical stimulation over the same area using microinjection of D,L-homocysteic acid (DLH) to preferentially activate somatodendritic receptors decreased the frequency of micturition but did not disrupt the coordinated pattern of voiding. In contrast, chemical stimulation within the caudal ventrolateral periaqueductal grey, in the area where critical synapses in the micturition reflex pathway are located, increased the frequency of micturition. CONCLUSION: Electrical deep brain stimulation within the midbrain can inhibit reflex micturition. We suggest that the applied stimulus entrained activity in the neural circuitry locally, thereby imposing an unphysiological pattern of activity. In a way similar to the use of electrical signals to 'jam' radio transmission, this may prevent a synchronized pattern of efferent activity being transmitted to the spinal outflows to orchestrate a coordinated voiding response. Further experiments to record neuronal firing in the midbrain during the deep brain stimulation will be necessary to test this hypothesis.

Myths and realities of the cardiac vagus.

There is continuing belief that cardiac parasympathetic postganglionic fibres are sparse or absent from the ventricles. This review of the literature shows that the supposition is a myth. Early studies considered that fine silver-stained fibres coursing amongst ventricle myocardial cells were most likely cardiac parasympathetic postganglionic fibres. The conclusions were later supported by acetyl cholinesterase staining using a method that appeared not to be associated with noradrenaline nerve fibres. The conclusion is critically examined in the light of several recent histological studies using the acetyl cholinesterase method and also a more definitive technique (CHAT), that suggest a widespread location of parasympathetic ganglia and a relatively dense parasympathetic innervation of ventricular muscle in a range of mammals including man. The many studies demonstrating acetylcholine release in the ventricle on vagal nerve stimulation and a high density of acetylcholine M2 receptors is in accord with this as are tests of ventricular performance from many physiological studies. Selective control of cardiac functions by anatomically segregated parasympathetic ganglia is discussed. It is argued that the influence of vagal stimulation on ventricular myocardial action potential refractory period, duration, force and rhythm is evidence that vagal fibres have close apposition to myocardial fibres. This is supported by clear evidence of accentuated antagonism between sympathetic activity and vagal activity in the ventricle and also by direct effects of vagal activity independent of sympathetic activity. The idea of differential control of atrial and ventricular physiology by vagal C and vagal B preganglionic fibres is examined as well as differences in chemical phenotypes and their function. The latter is reflected in medullary and supramedullary control. Reference is made to the importance of this knowledge to understanding the normal physiology of cardiac autonomic control and significance to pathology.

The projection and synaptic organisation of NTS afferent connections with presympathetic neurons, GABA and nNOS neurons in the paraventricular nucleus of the hypothalamus.

Elevated sympathetic nerve activity, strongly associated with cardiovascular disease, is partly generated from the presympathetic neurons of the paraventricular nucleus of the hypothalamus (PVN). The PVN-presympathetic neurons regulating cardiac and vasomotor sympathetic activity receive information about cardiovascular status from receptors in the heart and circulation. These receptors signal changes via afferent neurons terminating in the nucleus tractus solitarius (NTS), some of which may result in excitation or inhibition of PVN-presympathetic neurons. Understanding the anatomy and neurochemistry of NTS afferent connections within the PVN could provide important clues to the impairment in homeostasis cardiovascular control associated with disease. Transynaptic labelling has shown the presence of neuronal nitric oxide synthase (nNOS)-containing neurons and GABA interneurons that terminate on presympathetic PVN neurons any of which may be the target for NTS afferents. So far NTS connections to these diverse neuronal pools have not been demonstrated and were investigated in this study. Anterograde (biotin dextran amine - BDA) labelling of the ascending projection from the NTS and retrograde (fluorogold - FG or cholera toxin B subunit - CTB) labelling of PVN presympathetic neurons combined with immunohistochemistry for GABA and nNOS was used to identify the terminal neuronal targets of the ascending projection from the NTS. It was shown that NTS afferent terminals are apposed to either PVN-GABA interneurons or to nitric oxide producing neurons or even directly to presympathetic neurons. Furthermore, there was evidence that some NTS axons were positive for vesicular glutamate transporter 2 (vGLUT2). The data provide an anatomical basis for the different functions of cardiovascular receptors that mediate their actions via the NTS-PVN pathways.

GABAergic control of micturition within the periaqueductal grey matter of the male rat.

In urethane-anaesthetised rats continuous infusion of saline into the bladder (6 ml h⁻¹) evoked periodic sharp rises in intravesicular pressure accompanied by rhythmic bursting of external urethral sphincter (EUS) EMG and expulsion of urine from the urethral meatus. Microinjection of the GABA agonist muscimol (250 pmol) into the caudal ventrolateral periaqueductal grey (PAG), but not at other sites in the PAG, either depressed reflex voiding frequency (-60%, n = 7) and tonic EUS EMG activity (-38%, n = 6) or completely inhibited voiding (four sites). Microinjection of the GABA antagonist bicuculline (BIC; 1 nmol) into the same region, to reduce ongoing GABA tone, increased reflex voiding frequency (+467%, n = 16) and tonic activity in the EUS (+56%, n = 7) whilst bursting activity in the EUS became desynchronised. Although muscimol failed to change reflex micturition when microinjected into the dorsal caudal PAG, microinjection of BIC at these sites evoked pronounced autonomic arousal and increased reflex voiding frequency (+237%, n = 34). The results demonstrate that the functional integrity of synapses in the caudal ventrolateral PAG is essential to permit micturition. Transmission through the region is normally regulated by a tonic GABAergic inhibitory influence. In contrast, the functional integrity of the dorsal caudal PAG is not essential for reflex micturition. However, micturition may be initiated from this region via projections to the caudal ventrolateral PAG, as part of the behavioural response to psychological threat or other stressful stimuli.

Haemolytic and cytotoxic activities of the Tween 80-extracted putative haemolysin of Pasteurella multocida B:2.

The objective of this study was to investigate the haemolytic and cytotoxic activity of Pasteurella multocida B:2 strains, originally from cases of haemorrhagic septicaemia in cattle. All six P. multocida B:2 strains were non-haemolytic on sheep blood agar (SBA) and horse blood agar (HBA) when grown aerobically and on SBA anaerobically but they were haemolytic on HBA when grown anaerobically. No haemolytic activity against horse red blood cells was detected in culture supernates from aerobically or anaerobically grown cultures and only very weak haemolytic activity was obtained in supernates or pellet fractions from sonicated cells. However, after repeated extraction of sonicated cells with Tween 80, haemolytic activity was found in various cell fractions, both Tween-soluble and -insoluble. The Tween-extracted putative haemolysin and other bacterial fractions were also cytotoxic for mouse macrophage-like J774.2 cells. Further characterisation of the putative haemolysin revealed it to be a heat-labile, non-pore-forming protein of molecular weight >10 kDa whose activity was completely destroyed by trypsin and greatly reduced with protease and proteinase K treatment. Congo red also reduced the haemolytic activity. Non-denaturing gel-electrophoresis and RBC agar overlay revealed clear haemolytic zones but suggested that Tween was bound to some component of the P. multocida B:2 fractions and was responsible, to some extent, for the haemolytic activity observed. However, the effect of heat and other reagents on the Tween-extracted fractions and the lack of haemolytic activity in different Tween-extracted cell fractions of organisms other than P. multocida suggested that some proteinaceous component of the organism could indeed act as a haemolysin. This putative haemolysin may be one of the virulence attributes of P. multocida, but its characterisation and role in pathogenesis require further study.

Virulence spectra of typed strains of Campylobacter jejuni from different sources: a blinded in vivo study.

Campylobacter jejuni is a major cause of human diarrhoeal disease, but specific virulence mechanisms have not been well defined. The aims of the present blinded study were to measure and compare the in vivo properties of 40 serotyped, biotyped and genotyped C. jejuni isolates from different sources and genetic makeup. An 11-day-old chick embryo lethality assay, which measured embryo deaths and total viable bacteria over 72 h following inoculation of bacteria into the chorioallantoic membrane, revealed a spectrum of activity within the C. jejuni strains. Human and chicken isolates showed similar high virulence values for embryo deaths while the virulence of the bovine isolates was less pronounced. A one-way ANOVA comparison between the capacity of the strains to kill the chick embryos after 24 h with cytotoxicity towards cultured CaCo-2 cells was significant (P=0.025). After inoculation with a Campylobacter strain, mouse ligated ileal loops were examined histologically and revealed degrees of villous atrophy, abnormal mucosa, dilation of the lumen, congestion and blood in lumen, depending on the isolate examined. A 'total pathology score', derived for each C. jejuni strain after grading the pathology features for degree of severity, showed no apparent relationship with the source of isolation. Some relationship was found between amplified fragment length polymorphism groups and total ileal loop pathology scores, and a one-way ANOVA comparison of the mouse pathology scores against total chick embryo deaths after 72 h was significant (P=0.049).

Role of supraspinal vasopressin neurones in the effects of atrial natriuretic peptide on sympathetic nerve activity.

The aim of the present study was to determine if paraventricular-spinal vasopressin neurones participate in the sympatho-inhibitory effects of systemically administered atrial natriuretic peptide (ANP) on renal sympathetic nerve activity (RSNA). Experiments were carried out on male Sprague-Dawley rats anesthetized with 1.3 g/kg urethane. Changes in mean arterial pressure (mm Hg), heart rate (beats per minute) and RSNA (%) were measured following intravenous bolus administration of ANP (250 ng, 500 ng and 5 microg). Intrathecal application of selective V 1a receptor antagonist was performed to test for the involvement of supraspinal vasopressin pathways in mediating the effect on sympathetic outflow evoked by intravenous ANP administration. The results obtained demonstrated that both low and high doses of ANP caused renal sympathoinhibition (250 ng; - 7.5 +/- 1%, 500 ng; - 14.2 +/- 1%, 5 microg; - 16.4 +/- 2%), concomitant with vasodilation and bradycardia. After spinal vasopressin receptor blockade, the inhibitory effects of ANP were prevented and there was a small renal sympatho-excitation (250 ng; + 1.7 +/- 0.2%, 500 ng; + 6.1 +/- 0.03%, 5 microg; + 8.0 +/- 0.03%, P < 0.05). Therefore, the renal sympathetic nerve inhibition elicited by circulating ANP is dependent on the efficacy of a well established supraspinal vasopressin pathway. Since supraspinal vasopressin neurones without exception excite renal sympathetic neurones, it is suggested that ANP elicits this effect by activating cardiac vagal afferents that inhibit the spinally projecting vasopressin neurones at their origin in the paraventricular nucleus of the hypothalamus.

Cardiac sympatho-excitatory action of PVN-spinal oxytocin neurones.

A significant proportion of the spinally projecting neurones in the paraventricular nucleus are immunoreactive for oxytocin. Some of these oxytocin neurones terminate on sympathetic preganglionic neurones in the upper thoracic spinal cord, a region from which cardiac sympathetic neurones originate. No studies have so far identified a cardiac action of the supraspinal oxytocin neurones. The present study was designed to test the hypothesis that these oxytocin neurones excite spinal cardiac sympathetic neurones. This was done by measuring heart rate changes in response to intrathecal oxytocin and a selective agonist, and to stimulation of paraventricular neurones before and during blockade of spinal sites with selective antagonists. Rats were anaesthetised with chloralose and urethane (50 mg and 650 mg/kg) and recordings were made of heart rate and blood pressure. Drugs in a volume of 10 microl were applied to the upper thoracic spinal cord via a catheter placed intrathecally with its tip at T2. The paraventricular nucleus was explored with a glass micropipette, placed stereotaxically, and filled with d,l-homocysteic acid (DLH, 200 mM) for exciting neurones and pontamine sky blue for marking the position. Oxytocin (0.002 mM) applied to the spinal cord elicited increases in heart rate (26+/-5 beats per minute). This was mimicked by a highly selective oxytocin agonist. These heart rate increases were blocked selectively by two different oxytocin antagonists but unaffected by a V(1a) vasopressin antagonist. Excitation of sites in dorsal and medial parvocellular sub-nuclei of the paraventricular nucleus elicited increases in heart rate (36+/-3 bpm) which were significantly reduced by oxytocin antagonists but not affected by V(1a) antagonist. Also these induced increases in heart rate were unaffected by vagotomy or i.v. atropine but were abolished by i.v. esmolol. It is concluded that there is a population of paraventricular-spinal oxytocin neurones that excite cardiac sympathetic preganglionic neurones controlling heart rate.

Electrophysiological characteristics of vasomotor preganglionic neurons and related neurons in the thoracic spinal cord of the rat: an intracellular study in vivo.

Sympathetic preganglionic neurons (SPN) represent the final central neurons in the sympathetic pathways which regulate vasomotor tone; they therefore play a pivotal role in the re-distribution of cardiac output to different vascular beds in response to environmental challenges. While the consensus view is that activity in these neurons is due mainly to supraspinal inputs, the possibility that some activity may be generated intrinsically and modified by synaptic inputs cannot be excluded. Therefore, in order to distinguish between these two possibilities, the electrophysiological properties of cardiovascular-like SPN in the upper thoracic spinal cord of the anesthetized rat were examined and their response to activation of vasodepressor inputs was investigated. Intracellular recordings were made from 22 antidromically identified SPN of which 17 displayed irregular, but maintained, spontaneous activity; no evidence of bursting behavior or pacemaker-like activity was observed. Stimulation of the aortic depressor nerve or a vasodepressor site within the nucleus tractus solitarius (NTS) resulted in a membrane hyperpolarization, decrease in cell input resistance and long-lasting cessation of neuronal firing in SPN including a sub-population which had cardiac-modulated patterns of activity patterns. Recordings were also undertaken from 80 non-antidromically-activated neurons located in the vicinity of SPN; 23% of which fired in phase with the cardiac cycle, with this peak of activity occurring before similar increases in cardiac-modulated SPN. Stimulation of vasodepressor regions of the NTS evoked a membrane hyperpolarization and decrease in cell input resistance in cardiac-modulated but not non-modulated interneurons. These studies show that activity patterns in SPN in vivo are determined principally by synaptic inputs. They also demonstrate that spinal interneurons which exhibit cardiac-modulated patterns of activity are postsynaptically inhibited following activation of baroreceptor pathways. However, the question as to whether these inhibitory pathways and/or disfacilitation of tonic excitatory drive underlies the baroreceptor-mediated inhibition of SPN remains to be determined.

Elevated expression of adenosine A1 receptor in bronchial biopsy specimens from asthmatic subjects.

Asthmatics, unlike healthy subjects, experience bronchoconstriction in response to inhaled adenosine, and extracellular adenosine concentrations are elevated in the bronchoalveolar lavage fluid and exhaled breath condensate of asthmatic subjects. However, little is known about the location and expression of adenosine receptors in asthmatic airways. The aim of the present study was to investigate the distribution of adenosine A(1) receptors in bronchial biopsy specimens from mildly asthmatic steroid-naïve subjects and then compare the degree of expression with that of healthy subjects. Biopsy sections were immunostained using an adenosine A(1) receptor antibody, the selectivity of which was validated in specific experiments. Image analysis was then performed in order to determine differences in immunostaining intensity. Immunostaining of biopsy sections from the asthmatic subjects revealed strong expression of the A(1) receptor, located predominantly in the bronchial epithelium and bronchial smooth muscle. In comparison, very weak immunostaining was observed in biopsy specimens obtained from healthy subjects. Image analysis revealed that the intensity of positive staining of the asthmatic bronchial epithelium and smooth muscle regions was significantly greater than that observed for the healthy epithelium and smooth muscle. In conclusion, the sensitivity of asthmatics to inhaled adenosine coupled with increased adenosine A(1) receptor expression implies that these receptors play a role in the pathophysiology of this disease.

A physiological role for nitric oxide in the centrally mediated sympathetic and somatomotor ejaculatory response in anesthetized male Wistar rats.

Neurones in the lumbosacral spinal cord are known to play a significant role in ejaculation. In these same areas neurones containing nitric oxide synthase (NOS) have been described. This raised the question as to whether there is a physiological role for nitrous oxide (NO) in the spinal cord in sexual behavior. We first established immunohistochemical localization of NOS positive neurones in the lumbosacral spinal cord. NOS positive neurones were found in several areas of the lumbosacral cord. Namely the intermediolateral column (IML) at L(1)-L(4) and sacral cord; the dorsal gray commissure above the central canal at L(1)-L(2); the ventral gray area of lamina X below the central canal at L(3)-L(4); the superficial laminae of the dorsal horn at all levels. Secondly, we examined the role of NO in the generation of synchronized bursting activity within the vas deferens nerve and associated penile muscles, typical of sexual responses in the male anesthetized rat. NO modulators were applied directly to the spinal cord at T(13)-L(4) via a catheter placed subdurally (intrathecal) and their effect on the genital responses evoked by systemic administration of p-chloroamphetamine (PCA) or apomorphine (apo) (both 1 mg/kg) was observed. All responses evoked by PCA (n=4) or apo (n=3) were abolished or reduced (n=1) during intrathecal NOS inhibition using N((G)) nitro-L-Arginine methyl ester (l-NAME, 200 mM, 20-microl). NOS inhibition using l-NAME was reversed with simultaneous intrathecal application of the NO substrate l-arginine (100 mM, 20-microl, n=3). The selective neuronal NOS inhibitor 1-(2-trifluoro-methylphenyl) imidazole (100 mM, 20-microl, TRIM) also abolished all responses evoked by PCA (n=3). There was evidence that the responses within the vas deferens nerve (VDN) after PCA or apo were enhanced following NO donation using sodium nitroprusside (SNP, 1 mM, 20-microl). Furthermore, a PCA-like response within the VDN was evoked following intrathecally applied l-glutamic acid (200 mM, 20-microl) in six of 26 animals and also by intrathecal SNP in two of eight animals. In conclusion the results suggest a significant excitatory role for NO in the bursting pattern of synchronized discharge generated in autonomic and somatic outflows from the lumbosacral cord by neurones governing ejaculation.

Comparison of virulence-associated in vitro properties of typed strains of Campylobacter jejuni from different sources.

Campylobacter jejuni is a major cause of human diarrhoeal disease, but specific virulence mechanisms have not been well defined. This blinded study was undertaken with 40 C. jejuni isolates from different sources to determine their haemolytic, cytotoxic and adhesion and invasion activities towards mammalian cells. The results were correlated with source of isolation and genetic makeup by amplified fragment length polymorphism (AFLP) typing. The isolates had variable degrees of haemolytic activity against rabbit erythrocytes and cytotoxicity towards CaCo-2, HeLa and Vero cells. The data indicated that the haemolytic and cytotoxic activities were due to separate factors. A range of cytotoxicity was exhibited, whereby some strains had no activity against the target cells and others had activity against all three cell lines. Certain strains had activity against CaCo-2 cells but little or no activity against the other cells, while others exhibited the opposite phenotype. The data suggested that the cytotoxicity assay with the different cell lines may have detected more than one cytotoxin. A wide variation between isolates was observed for both adherence and invasion with all three cell lines, yet, overall, the strains showed a significantly greater invasion capacity for CaCo-2. There was no clear relationship between source of isolation or disease manifestation and possession of statistically significantly higher levels of particular virulence-associated factors although, in some cases, a correlation between cytotoxicity and cell invasion was evident. Five AFLP clusters, each representing two to eleven isolates with similar profiles, were observed at the 90 % similarity level. Some AFLP groups contained isolates with a common serotype, but each group had C. jejuni isolates from more than one source with the exception of group IV, which contained only human isolates. Isolates with high cytotoxic activity against CaCo-2 cells were confined to groups I, III and IV and a group of unrelated strains (U). Group II isolates had uniformly low cytotoxicity. Isolates in groups I, V and U were more invasive for CaCo-2 cells than isolates in groups II, III and IV. The strain differences in cytotoxicity or invasion did not correlate with source of isolation.

Pain and changes in peripheral resistance at high vascular transmural pressure in the human forearm.

A hydrostatic rise in forearm vascular transmural pressure may be associated with an increase in forearm blood flow (FBF) that causes pain. To test this hypothesis, forearm vascular transmural pressure was elevated in eight male volunteers by a series of 1-min hypobaric exposures of the left arm to incrementing differential pressures of 40, 80, 120, 140, 160 and 200 mmHg. The series was repeated after a 30-min interval. Forearm venous pressure (FVP) was measured in the median antecubital vein, and FBF was determined by ultrasound Doppler in the axillary artery. Pain level was recorded by numerical rating scale. In all subjects, an increase in FBF and forearm vascular conductance (FVC) occurred (P < 0.05) at high FVP (mean +/- SE, 184 +/- 8 mmHg). Pain was linearly related to the increase in FVC. In the second series of exposures, increases in FBF, FVC and pain occurred at a lower transmural pressure (FVP 152 +/- 15 mmHg, P < 0.01). It is concluded that intense forearm pain is associated with a failure of autoregulation in the peripheral vascular bed and is worsened on repeated exposure to high transmural pressure. This may explain the overt forearm pain experienced by the crews of high performance military aircraft during manoeuvring.

Failure of vascular autoregulation in the upper limb with increased +Gz acceleration.

Forearm pain occurring during high +Gz exposure has been linked with vascular distension from elevated transmural pressure of hydrostatic origin and is exacerbated by positive pressure breathing (PBG). We postulated that at high vascular transmural pressure vascular autoregulation might be overcome and be associated with worsened pain. Six volunteers were studied at +4, +5, +6, and +7 Gz on a human centrifuge. Forearm vascular resistance (FVR) was assessed by Doppler ultrasound resistive index (RI), and superficial forearm venous pressure (FVP) was measured via an indwelling catheter. Pain rating was assessed by numerical scale. The left arm was located at heart level (control position), or on the throttle (test position). Runs were completed with and without positive pressure breathing for G protection (PBG); subjects wore full coverage anti-G trousers and chest counter-pressure. In the test position, pain increased with increasing acceleration (P < 0.0001), and was more severe with PBG at +5 Gz and +7 Gz (P < 0.05). FVP rose substantially more in the test than control position (238 +/- 17 mmHg vs. 61 +/- 8 mmHg at +7 Gz, P < 0.0001) but the presence or absence of PBG had no effect on the FVP increase during acceleration in either position. In the test position, RI fell with increasing acceleration above +5 Gz (P < 0.0001), and the fall was greater with PBG (P < 0.05). Forearm pain was thus associated with a decrease in FVR and an increase in vascular transmural pressure. PBG exacerbated forearm pain and prompted a greater fall in RI, but had no effect on FVP response. These findings support FVR but not forearm venous distension in the aetiology of +Gz arm pain.