Kaposi sarcoma (KS) is a multifocal, vascular lesion of low-grade malignant potential that presents most frequently in mucocutaneous sites. KS also commonly involves lymph nodes and visceral organs. This article deals with the manifestation of KS in unusual anatomic regions. Unusual locations of KS involvement include the musculoskeletal system, central and peripheral nervous system, larynx, eye, major salivary glands, endocrine organs, heart, thoracic duct, urinary system and breast. The development of KS within wounds and blood clots is also presented. KS in these atypical sites may prove difficult to diagnose, resulting in patient mismanagement. Theories to explain the rarity and development of KS in these unusual sites are discussed. Kaposi sarcoma (KS) is a vascular lesion of low-grade malignant potential that is associated with Human Herpesvirus-8 (HHV8) infection. KS is a multifocal tumor that manifests most frequently in mucocutaneous sites, typically the skin of the lower extremities, face, trunk, genitalia and oropharyngeal mucosa Table . KS alsoKS involvement of the musculoskeletal system is a rare occurrence . NeverthThe musculoskeletal system has been shown to be involved by all KS epidemiological forms, including AIDS-related, Classic, African (endemic), and infrequently transplanted-associated KS ,18-21. Rrd, 4th and 5th metatarsals). Asymmetric bone involvement by KS appears to be the rule. Joint involvement is unusual, but has been reported [African and Classic KS lesions tend to involve the peripheral skeleton, whereas AIDS-related KS more commonly involves the axial skeleton  and/or maxillofacial bones ,23-32. Kreported .3[Osseous KS lesions are more likely to be associated with long-standing and locally aggressive cutaneous African and Classic KS lesions, or with AIDS-associated mucosal KS lesions that penetrate underlying bone . Patient3. AIDS-re3,38.Most osseous KS lesions are osteolytic, with cortical and at times almost complete bone destruction (osteolysis). KS of the bone does not present well on plain radiographs, despite their frequent osteolytic nature -40. CT sPeripheral nerve involvement by KS is rare. In a postmortem study of temporal bones obtained from patients with AIDS, investigators discovered KS in the eighth cranial nerve of one patient . An autoAround 15 cases of suspected brain involvement with KS have been reported -58, but A few of the cases reported have been regarded with some suspicion by others, as the patients in these reports did not have skin lesions . Based uThere have been two cases of KS infiltrating the spinal cord ,48. In oHead and neck involvement of KS is not unusual. However, laryngeal involvement is somewhat of an infrequent manifestation. There have been approximately 25 accounts of KS of the larynx -73. MostPresenting symptoms may include hoarseness, throat discomfort, urge to cough, aphonia, dysphagia, stridor or complete airway obstruction. Examination may reveal laryngeal edema or more likely a purple vascular mass lesion. The lesion's surface can appear verrucous in nature, due to deposits of dry secretion . The diaTherapeutic options for this scenario include low-dose local irradiation, intralesional chemotherapy or laser ablation, and systemic therapy, particularly if there is disseminated KS. For laryngeal KS lesions producing acute or impending airway obstruction urgent intervention is necessary. However, depending on the location of the KS lesion, tracheostomy may contribute to mortality as a result of fatal hemorrhage . TherefoPeriorbital edema may occur with KS of the face ,76. ExteExternal ocular KS may manifest as a mass lesion or simply as a subconjunctival hemorrhage. Interestingly, increased sludging of conjunctival blood-flow has been shown in patients with KS . EpiphorKS can involve the parenchyma and/or intraparenchymal lymph nodes of the major salivary glands . Both thVery few detailed case reports of adrenal KS have been published, mostly in patients with AIDS ,106. AdrInvolvement of the thyroid gland by KS is exceedingly rare, and presently only 5 cases have been reported ,107-111.No report of KS involving the pituitary was found. In particular, KS of the pituitary gland was not identified in a series of 49 autopsied patients with AIDS in which the pituitary gland was specifically studied .Cardiac involvement has been reported in the various epidemiologic forms of KS. The results of autopsy studies conducted from 1959–1986 identified KS of the heart in 18% of African (endemic), 17% in AIDS-related (epidemic), and 15% of Classic (sporadic) KS ,113. SurChylothorax is a known, but rare manifestation of KS involving the thoracic duct and adjacent mediastinal structures -123. KS Genital KS lesions are common. Infrequently, urethral meatal lesions may cause outlet obstruction and urinary retention ,126. HowSince HHV8 can be detected in breast milk , it is nLocalization of KS to sites of previous iatrogenic trauma has been documented ,139, sevA case of transplant-related KS restricted to the site of a previous deep venous thrombosis has been documented . This 72It is clear from the aforementioned review that KS has been reported in virtually all anatomic sites. KS involvement of other unusual sites not discussed in this chapter include the gonads (e.g. testis) , endometAs a result of HIV, a significant increase in anatomically disseminated KS cases was evident. The anatomical distribution of KS lesions also changed during the AIDS period. Soon after the onset of the AIDS epidemic, head and neck AIDS-related KS became one of the most common manifestations of AIDS . In a stThe occurrence of KS in any of these atypical sites may prove difficult to diagnose, particularly if patients are asymptomatic, lesions go unrecognized on routine imaging studies (e.g. osseous KS on plain x-ray films), and/or clinician's are unwary of their existence . AwareneIn several cases, there was limited available clinical data. Pathologic verification of a diagnosis of KS by means of biopsy and/or autopsy was not available in some cases. In their review of 66 cases of reported osseous KS, the authors noted that histological proof of actual bone involvement by KS was only obtained in 53% of cases . Very raIt is interesting to hypothesize why KS is rarely encountered in the aforementioned anatomic locations. Moreover, why KS involvement of sites like the bladder and breast are uncommon when HHV8 is shed in urine and breast milk, respectively, from infected patients is intriguing. A urinary protein called ANUP (antineoplastic urinary protein) has certainly been shown to inhibit KS growth in a murine model . ConversIt is also unclear if isolated KS lesions in some of these sites represent primary or disseminated disease. Some authors have considered the occurrence of KS in unusual locations to represent metastatic disease ,160. AltKS lesions develop as a result of the following combination of factors: HHV8, altered immunity (immunosuppression), and an inflammatory/angiogenic milieu. This may explain why, in the appropriate clinical setting, KS develops in traumatized tissue or pemphigus lesions . Some auThe authors declare that they have no competing interests.LP and BJD contributed equally to all aspects of this paper.The pre-publication history for this paper can be accessed here:


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Indocyanine green (ICG) is clinically approved for the determination of liver function, cardiac output and plasma volume. In this pilot study, ICG was used as photosensitizer in combination with a diode laser to treat AIDS-associated Kaposi's sarcoma (KS) in three patients. Directly and up to 50 min after intravenous administration of ICG (2-4 mg kg(-1) body weight), KS (n=57), mainly plaque-type, were irradiated using a diode laser  matching the absorption maximum. Complete remission of KS (n=16) was achieved when irradiated 1-30 min after injection of the second dose of ICG (2 x 2 mg kg(-1) b.w., 30 min apart) with 3-5 W cm(-2) and 100 J cm(-2). Biopsies (n=3) revealed necrosis of the tumour 24 h and complete remission 4 weeks after therapy. In general, systemic side-effects were not observed and cosmetic results were very good. However, hyperpigmentation occurred temporarily in lesions located on the lower extremities. These findings show that AIDS-associated KS can be effectively treated after photosensitization with ICG and subsequent irradiation with an appropriate diode laser. However, additional investigations need to elucidate the exact mechanism of action of ICG-mediated phototherapy and have to show the efficacy for the treatment of other highly vascularized solid tumours.


We used a plate immunoassay to monitor trafficking of these proteins.We measured activity of the dopamine transporter (DAT) by the efflux of −14 to 10−11 M DDE caused significant efflux inhibition, whereas NP and BPA enhanced or inhibited efflux at several concentrations. We also measured the effects of E2/XE combinations; DDE potentiated E2-mediated dopamine efflux, whereas BPA inhibited it. In E2-induced efflux, 15% more ER-α trafficked to the membrane, whereas ER-β waned; during BPA-induced efflux, 20% more DAT was trafficked to the plasma membrane.All compounds at 1 nM either caused efflux or inhibited efflux, or both; each compound evoked a distinct oscillatory pattern. At optimal times for each effect, we examined different concentrations of XEs. All XEs were active at some concentration < 10 nM, and dose responses were all nonmonotonic. For example, 10Low levels of environmental estrogen contaminants acting as endocrine disruptors via membrane ERs can alter dopamine efflux temporal patterning and the trafficking of DAT and membrane ERs, providing a cellular mechanism that could explain the disruption of physiologic neurotransmitter function. In addition, XEs break down slowly, so persistent deposits are found in the soil (Xenoestrogens (XEs) are environmental contaminants capable of mimicking the effects of endogenous estrogens, but usually not precisely. Thus, they can initiate more, different, and mistimed estrogen exposures that can lead to disruptions of estrogenic signaling . XEs canthe soil , thus ex2), estrone (E1), and estriol (E3)   can act pathways .2 can affect both the DAT (The dopamine transporter (DAT) is a selective transporter that primarily functions to stop dopamine signaling by removing dopamine (dopamine uptake) from the extracellular space after a stimulatory release of dopamine has occurred. Dopamine is subsequently brought into intracellular vesicles via vesicular monoamine transporters to protect it and maintain dopamine separation from other cellular molecules with which it might react. There it is stored for subsequent rapid signaling  resulted in decreased dopamine levels and impaired memory in Wistar rat pups . Polychlma cells . DDE, dima cells  in in vi2 inhibits dopamine uptake  can cause either potentiation or attenuation of E2-mediated dopamine efflux, depending on the estrogen mimetic coadministered. Finally, we report a functional link between the subcellular location of the ER subtypes and DAT resulting from BPA treatment compared with treatment with E2. These cellular events could explain the impact of XE exposures on dopamine efflux, including changes in neurotransmission, and some forms of functional endocrine disruption associated with neurotoxicity, especially in the context of diseases that are affected differentially between males and females.We recently used nerve growth factor (NGF)-differentiated PC12 cells, a well-characterized neuronal cell model, to demon strate that Ee uptake  and subse uptake . Here we3H-dopamine from PerkinElmer ; Scintiverse II from Fisher ; DDE and endosulfan from Ultra Scientific ; and ABC (avidin:biotinylated enzyme complex) with alkaline phosphatase, para-nitrophenol phosphate, and levamisole from Vector Laboratories . All other compounds, if not specified in text, were obtained from Sigma Chemical Co. .We purchased fetal bovine serum (FBS) from Atlanta Biologicals ; equine serum from Gibco ; PC12 cells were grown in high-glucose, phenol red–free RPMI 1640 medium containing 5% FBS and 5% equine serum. For all experiments, cells were transferred to medium supplemented with 0.5% 4× charcoal-stripped FBS and 0.5% equine serum 48 hr before experimentation to minimize the effects of endogenous hormones. To promote differentiation and up-regulate DAT and mER-α, 20 ng/mL NGF-β was also added for 48 hr. Cells of passages 61–64 were used for these studies.3H-dopamine efflux using selective inhibitors to define the transporter. We plated PC12 cells on poly--lysine (10 μg/mL)-coated 48-well plates, deprived of endogenous hormones. We added uptake buffer  containing 0.2 mg/mL ascorbic acid and 50 nM desipramine (pH 7.4) with or without 1-(2-[bis(4-fluorophenyl)methoxy]ethyl)-4-(3-phenylpropyl) piperazine dihydro chloride (GBR 1290) for 60 min at 37°C. GBR 12909 (100 nM) was added for a 60-min preincubation to define selective efflux by DAT. 3H-Dopamine (20 nM) was loaded into the cells for 10 min before two washes in release buffer . We then added release buffer containing treatments with or without GBR 12909 and collected extracellular fluid at the indicated time points to assess efflux. Triplicate aliquots of each well were counted in 2 mL Scintiverse II scintillant using a Beckman LS600SE scintillation counter  averaged to provide one sample (n = 1) per well. Specific efflux was defined by averaging the disintegrations per minute due to efflux in the presence of desipramine and GBR 12909, and then subtracting these values from the efflux observed with desipramine alone. We subtracted background (vehicle controls) matched for ethanol concentration and time from treatment groups and present the data as 3H-dopamine efflux per 106 cells. Our rationale for using 1 nM E2 in combination with a range of concentrations for the XEs DDE and BPA was the prevalence of approximately 1 nM E2 as a physiologic concentration of total circulating E2 to which XEs would likely be added.As described previously , we adappara-nitrophenol phosphate plus 0.5 mM levamisole was added in 100 mM sodium bicarbonate solution for 50 min at 37°C. Plates were read at 405 nm absorbance (A405), rinsed and stained with 0.1% crystal violet for 30 min at room temperature, washed with double-distilled H2O, and dried overnight. We read dye (extracted from each well with 50 μL 10% acetic acid) at A590 and used this to estimate cell number per well. Data are plotted as percentage of control levels (A405/A590).Briefly, PC12 cells were plated on poly--lysine (10 μg/mL)-coated 96-well plates at 5,000 cells/well, as previously described . Differep < 0.05.We repeated each experimental set three times on different cell preparations, and we report the number for each treatment in the figure legends. We used analysis of variance to indicate whether further testing of individual data points for significance from their control was warranted. We performed statistical analyses for all assays using SigmaStat software  and set statistical significance at −9 M E2 causes rapid dopamine efflux of preloaded 3H-dopamine from PC12 cells (2 compared with each XE that we studied at this concentration. Both E2 and diethyl stilbestrol (DES) displayed peak dopamine efflux at 9–15 min, followed by a return to baseline. NP and BPA caused rapid dopamine efflux within 5 min, followed by a return to baseline for NP and inhibited dopamine efflux for BPA. The pesticides DDE, dieldrin, and endosulfan did not cause dopamine efflux, but dopamine efflux inhibition occurred as early as 1 min for dieldrin and 3 min for DDE and endo-sulfan, followed by continuous baseline activity. Interestingly, our results suggest that the compounds that belong to the same functional use/chemical structure class  cause similar temporal and stimulatory/inhibitory regulation of dopamine efflux. The rapidity with which all tested compounds in these studies either cause or inhibit dopamine efflux implies that nongenomic actions are involved.We have previously demonstrated that 1012 cells . Figure 2 caused dopamine efflux at the lowest concentration tested (10−14M) and at the higher but still physiologic concentrations (10−10 to 10−8 M), but not at the intermediate concentrations  at times and concentrations representing their maximum effects on efflux enhancement or inhibition. We assessed the membrane versus total cellular levels of these proteins in our quantitative immunoreactive plate assay (−9 M E2 treatment increased membrane levels of ER-α (a stimulator of efflux at this time and concentration) and decreased membrane ER-β (an inhibitor of this response), whereas total levels of these proteins declined in the cells  compared with either compound alone  in the presence of physiologic levels of E2, which was not significantly different from BPA or E2 alone at that concentration. Therefore, these two different XEs in combination with E2 altered dopamine efflux states compared with their singular effects in dramatically different ways.In our natural environment, exposure to these compounds is combinatorial, due to not only the presence of multiple compounds simultaneously but also the presence of endogenous steroids. Therefore, we examined combinations for a limited number of these cases by comparing single exposures with a physiologic dose of Ewith DDE  and BPA with DDE  at multind alone . The comalone  caused rapid dopamine efflux, similarly peaking at 9–15 min. DES also has genomic actions similar to those of E2  binding to ER-β and GPR30 in our cell model could affect estrogen-mediated dopamine efflux.On the basis of the dopamine efflux responses to these different estrogens (time courses and concentrations curves), we observed some similar patterns within subclasses of these chemicals. First, the group consisting of Ese of E2 . However studies . Howeverudies . This (and other examples from this series of experiments) demonstrates that XEs, causing efflux in distinctly different temporal patterns compared with E2, could cause altered patterns of response in combination. We have previously reported that some XEs also elicit responses differentially in pituitary cells , which is oxidized to MPP+ (1-methyl-4-phenyl-pyridinium), the primary constituent of the herbicide cyperquat . They caused rapid inhibition of dopamine efflux at the lower concentrations. Some organochlorine pesticides, such as DDE and dieldrin, have been shown to cause increased dopamine, DAT, and vesicular monoamine transporter levels in vivo . In uteryperquat . These eyperquat , suggest2 and all XEs that we studied can cause dopamine efflux changes at low, environmentally relevant concentrations. We also show that at some of these rapid time points and low concentrations, these compounds caused significant inhibition of dopamine efflux. Only a careful mapping of the response effects of combinations of these and other estrogenic compounds  will fully elucidate their aggregate potential for influencing dopamine efflux and thus contributing to adverse neurologic outcomes.Our extensive range of dose–response curves, at two time points for some compounds, revealed that E2. Recently, in vitro  matched human postmortem levels in Parkinson disease patients; these levels inhibit dopamine uptake and cause synaptosomal and vesicular release of dopamine without increasing intracellular oxidative stress. Increasing intra-cellular dopamine concentrations (by depleting vesicular stores) can promote dopamine efflux by exchange diffusion (2 we are increasing intra cellular dopamine to the point that the mechanism shifts from efflux inhibition (or uptake) to efflux. This is only one of many possible mechanisms that should be explored for combinations of XEs and physiologic estrogens, or other XEs.Although DDE alone did not cause dopamine efflux, it caused potentiation of dopamine efflux in the presence of physiologic levels of Eiffusion . It is p2. Compounds such as BPA that increase DAT numbers at the membrane have been reported to be pathological because increased DATs in the membrane leads to increased toxin susceptibility  is reported to be neuroprotective  leave the plasma membrane. The increased levels of membrane DAT caused by efflux-producing BPA treatments suggest that BPA-mediated dopamine efflux involves distinctly different intermediary mechanisms compared with Etibility . Althougotective . Therefo2 levels (Women are abusing stimulants, such as amphetamine, methamphetamine, and cocaine, at an increasing rate. Although drug addiction and abuse rates involve many social factors, females display an association between sex steroid levels and drug behavioral and neurochemical effects . Ampheta2 levels . If XEs 2 levels  and morp2 levels . It rema


EHP as we have focused on rebuilding the journal and its outreach programs. We appreciate the strong support you, our readers, have shown us. We, in turn, are committed to making EHP as strong as it can be, with new program features to carry our content further than ever.The past year has been an important time for Environ Health Perspect 116:1449–1455 (2008)]. We are also putting the EHP Science Education Program on a brief sabbatical while we conduct a thorough evaluation to guide us in best meeting the needs of our users. We will not be publishing new lessons during this period, but instead will be busy conducting focus groups, surveys, and resource needs analyses, all with an eye toward broadening the impact of the program both domestically and internationally. Finally, in summer 2009 we will debut the all-new EHP website (www.ehponline.org). The site will be packed with new features, including enhanced search capabilities and a “Comment” feature that will let you discuss news articles online.This spring we are pleased to announce the launch of our new podcast series, “The Researcher’s Perspective.” Our inaugural podcast is an interview with Kristie Ebi, who discusses her recent paper “Climate Change, Tropospheric Ozone and Particulate Matter, and Health Impacts” [As always, we welcome your feedback and thank you for your support.


Jacob, a recently identified Caldendrin binding partner, is a novel protein abundantly expressed in limbic brain and cerebral cortex. Strictly depending upon activation of NMDA-type glutamate receptors, Jacob is recruited to neuronal nuclei, resulting in a rapid stripping of synaptic contacts and in a drastically altered morphology of the dendritic tree. Jacob's nuclear trafficking from distal dendrites crucially requires the classical Importin pathway. Caldendrin binds to Jacob's nuclear localization signal in a Ca2+-dependent manner, thereby controlling Jacob's extranuclear localization by competing with the binding of Importin-α to Jacob's nuclear localization signal. This competition requires sustained synapto-dendritic Ca2+ levels, which presumably cannot be achieved by activation of extrasynaptic NMDA receptors, but are confined to Ca2+ microdomains such as postsynaptic spines. Extrasynaptic NMDA receptors, as opposed to their synaptic counterparts, trigger the cAMP response element-binding protein (CREB) shut-off pathway, and cell death. We found that nuclear knockdown of Jacob prevents CREB shut-off after extrasynaptic NMDA receptor activation, whereas its nuclear overexpression induces CREB shut-off without NMDA receptor stimulation. Importantly, nuclear knockdown of Jacob attenuates NMDA-induced loss of synaptic contacts, and neuronal degeneration. This defines a novel mechanism of synapse-to-nucleus communication via a synaptic Ca2+-sensor protein, which links the activity of NMDA receptors to nuclear signalling events involved in modelling synapto-dendritic input and NMDA receptor–induced cellular degeneration.NMDA (N-methyl-D-aspartate) receptors and calcium can exert multiple and very divergent effects within neuronal cells, thereby impacting opposing occurrences such as synaptic plasticity and neuronal degeneration. The neuronal Ca Long-lasting changes in communication between nerve cells require the regulation of gene expression. The influx of calcium ions into the cell, particularly through membrane protein called NMDA receptors, plays a crucial role in this process by determining the type of gene expression induced. NMDA receptors can exert multiple and very divergent effects within neuronal cells by impacting opposing phenomena such as synaptic plasticity and neuronal degeneration. We identified a protein termed Jacob that appears to play a pivotal role in such processes by entering the nucleus in response to NMDA receptor activation and controlling gene expression that governs cell survival and the stability of synaptic cell contacts. Removal of Jacob from the nucleus protects neurons from NMDA receptor–induced cell death and increases phosphorylation of the transcription factor CREB, whereas the opposite occurs after targeting Jacob exclusively to the nucleus. The work defines a novel pathway of synapse-to-nucleus communication involved in modelling synapto-dendritic input and NMDA receptor–induced cellular degeneration. A new signaling mechanism from NMDA receptors to the nucleus plays an important role in the phosphorylation of the transcription factor CREB and neuronal cell survival. On the contrary, NMDA receptors are also important players in neurodegenerative processes. Although both aspects require gene expression, our knowledge is still sparse concerning how these fundamental processes are regulated at the molecular level. The Janus face of neuronal NMDA receptor signalling is probably best reflected by the fact that the influx of Ca2+ ions is thought to act as one of the major mediators of synapto-nuclear signalling [2+ microdomains coupled to the activation of synaptic and extrasynaptic NMDA receptors, and transducing incoming Ca2+ events to different downstream pathways [2+ influx through synaptic NMDA receptors trigger nuclear cAMP response element-binding protein (CREB) phosphorylation via an extracellular signal-regulated kinase (ERK)-dependent pathway, whereas Ca2+ influx through extrasynaptic NMDA receptors leads via an ERK-independent pathway to a dephosphorylation of CREB termed CREB shut-off. As opposed to the synaptic pathway, the CREB shut-off signal is coupled to neuronal degeneration and cell death [Cagnalling ,2 and ofgnalling . Within pathways –4. In a pathways –7 providll death –3,8–10.2+ exerts its signalling functions via a variety of Ca2+ sensor proteins, pathways that result in a nuclear response to synaptic activity have primarily been based on signalling via calmodulin (CaM) [2+-bound state, CaM alters the properties of several other proteins and signalling cascades that have been implicated in diverse neuronal functions [2+ ions, a variety of additional EF-hand proteins have been identified in neurons, termed neuronal Ca2+ sensor (NCS) proteins. These NCS are believed to serve more-specific functions in neurons [Although Cain (CaM) ,2. In itunctions ,12. Wher neurons –15.One of these NCS proteins is Caldendrin  ,17, a biThis strategy disclosed a novel Caldendrin-binding partner, named Jacob, which exhibits a remarkably restricted expression in cortical and limbic brain regions of mammals. We report that Jacob displays a distribution similar to that of Caldendrin in the PSD, dendritic spines, and dendrites, but in contrast to Caldendrin, is also found in neuronal nuclei. Activation of NMDA receptors induces nuclear trafficking of Jacob that is under the control of Caldendrin and Importin-α. Our data imply that nuclear Jacob participates in the CREB shut-off pathway, which might play a physiological as well as pathophysiological role in the control of dendritic cytoarchitecture, synapse number, and neuronal survival under conditions of increased NMDA receptor activity.Utilizing yeast two-hybrid screening to identify binding partners for Caldendrin, we obtained eight independent clones of a hitherto uncharacterized gene product, which we termed Jacob. The longest cDNA clone encompassed an open reading frame of 1,596 bp encoding a 532–amino acid (aa) protein with a calculated molecular weight of 60 kDa  and S1A.Analysis of the primary structure of Jacob revealed a putative N-terminal myristoylation site and several potential phosphorylation sites for protein kinase C (PKC), cAMP-/cGMP-dependent protein kinases, and protein tyrosine kinases  and S1A.In situ hybridization experiments revealed a strikingly restricted localization of Jacob transcripts in the limbic brain and cortical areas (unpublished data), showing extensive overlap with Caldendrin mRNA expression (see ). In accAt the ultrastructural level, Jacob IR was localized to a subset of asymmetric type I synapses on dendrites of cortical neurons F–2H. ApaThe architecture of the nucleus includes two overlapping nucleic acid–containing structures that are directly associated with the regulation of gene expression: the chromatin and the nuclear matrix. Therefore, we isolated highly pure nuclear matrix, heterochromatin, and euchromatin fractions. Interestingly, after chromatin fractionation, Jacob was found to be exclusively associated with the RNA polymerase II–containing euchromatin C. Moreov3H-myristic acid with a wild-type (WT)-Jacob-GFP construct, subsequent immunoprecipitation with a monoclonal anti-GFP antibody and immunoblotting revealed incorporation of radioactivity at a band immunoreactive to both a polyclonal GFP antibody  lacking the six basic amino acid residues between 247–252. Transfection of this construct in COS-7 cells resulted in an extranuclear localization of the mutant protein  or WT-Jacob constructs. Transfection of these different mutants had drastic effects on cell morphology. WT-Jacob-GFP–transfected neurons as compared to GFP controls exhibited more, but less-complex, dendritic processes . Quantitative immunoblot analysis and immunostainings showed that viral infection of cortical primary cultures led to a specific reduction of these nuclear Jacob isoforms , providing further evidence that the reduction of synaptic contacts directly correlates with the presence of Jacob in the nucleus.To elucidate functional consequences of nuclear versus extranuclear localization of Jacob in terms of structural plasticity, we transfected hippocampal primary neurons with different mutant (Δrocesses C and S3A to 24 h A–4C. In  to 24 h A. Intere to 24 h D and 4E.sfection E. This sex9-Jacob that contain the NLS but lack large parts of the carboxy-terminus (Myr-Δex9-Jacob-GFP), which accumulated in the nucleus after transfection of primary neurons, and its overexpression resulted in a comparable reduction of dendritic complexity and synapse number as those seen with ΔMyr-Jacob-GFP (C-term-Jacob-GFP) had no morphological consequences despite the presence of the NLS and its exclusive nuclear localization -Caldendrin fusion protein. In this assay, the interaction of Caldendrin and Jacob was found to be Cant Jacob B. Strikin tested C, and Can tested D. Furthen tested E. These n tested B. Importn buffer F. Thus, 2+ binding states of the EF-hands, which are not rigid structural units, but may function as hinges at low Ca2+ concentration [2+, which in turn results in different binding dynamics at the first hydrophobic pocket compared to CaM.Three-dimensional modelling substantiated the idea that structural differences in Jacob binding surfaces of Caldendrin and CaM  can explntration . Indeed,RGyaeR, and residues 260–269, IQrnfRkhlr) within a central region (residues 229–272) containing the bipartite NLS . We next investigated whether the binding of Importin-α1 can be competed by equimolar amounts of recombinant Caldendrin. Indeed, these studies revealed a competition between Caldendrin and Importin-α1 for binding to Jacob in the presence of Ca2+ , we analyzed the transport process of Jacob in more detail using confocal laser scan microscopy. A brief depolarization of hippocampal neurons with KCl also induced a translocation of Jacob and Importin-α1 to the nucleus in the presence of anisomycin  16, indicating that the interaction with Caldendrin masks the bipartite NLS of Jacob A and 8B.2+- and NMDA receptor–activated signalling pathways to the nucleus in neurons funnel through the activation of the transcription factor CREB [Myr-Jacob-GFP construct significantly reduced the levels of pCREB in hippocampal primary neurons as compared to untransfected or GFP-transfected controls under resting conditions . Therefore, it is conceivable that Jacob is indirectly coupled via CREB-binding proteins to the CREB signalosome. To further support a role in gene expression, we provided substantial evidence that Jacob is highly enriched in two nuclear compartments associated with gene transcription and pre-mRNA processing. Jacob is abundant in euchromatin fractions and therefore present at active sites of gene transcription. The protein harbours long stretches of basic amino acid residues, which are well suited for DNA binding, although no known DNA binding motif was identified in its primary structure. Particularly with regard to the phenotype of its nuclear overexpression that involves a rapid destabilization of synaptic contacts and a retraction of dendrites, and which cannot be explained entirely by CREB shut-off, it is reasonable to assume that Jacob will be part of additional nuclear signalling events.The nature of such signalling events will be obviously related to the circumstances of Jacob's nuclear trafficking. CREB shut-off has been largely assigned so far to pathophysiological insults, including spill-over of glutamate after excessive stimulation or reversal of glutamate transporters in the context of epileptic seizures or brain ischemia . This vi2+ sensor protein regulates the nuclear localization of a protein by competitive binding to its NLS in a Ca2+-dependent manner. The significance of this novel mechanism of neuronal Ca2+ signalling is further underscored by the fact that binding of Caldendrin is specific in that its ancestor and closest relative in brain, CaM, did not bind to Jacob at any Ca2+ concentration tested. This is of importance since CaM levels are probably more than a magnitude higher in neurons than those of Caldendrin [2+ binding affinities are comparable between both proteins [A further intriguing aspect of this study is that it provides the first demonstration that an EF-hand CaM-like Caldendrin , and Ca2proteins . Computeproteins  are coveproteins . Accordiproteins . The priproteins . Thus, t2+ binding proteins relate to Caldendrin's neuronal function? In contrast to the interaction with Jacob, Caldendrin binding to most of its interaction partners is Ca2+ independent, as already described above for the LC3 interaction [2+-, CaM-, and ATP-independent interaction of the C-terminal half of Caldendrin/CaBP1 was demonstrated for the inositol trisphosphate receptor ((InsP3R) [3-triggered intracellular Ca2+ release [2+-independent binding was reported for L-type voltage-dependent CaV1.2 Ca2+ channels [2+ currents following synaptic activation and thereby indirectly via increased synaptic activity could promote Caldendrin's and possibly Jacob's synaptic localization. Low synaptic activity and, hence, low synapto-dendritic Ca2+ levels will instead favour Caldendrin's binding to the InsP3R. It is therefore conceivable that Caldendrin can thereby directly lower Ca2+ levels in dendritic microdomains, and in consequence, negatively regulate its own association with Jacob. Therefore, a switch of binding partners could directly relate to Caldendrin's role in regulating Jacob's nuclear transition. Along these lines, it can be predicted that keeping the delicate balance between Jacob's nuclear and extranuclear localization via Caldendrin binding will provide a powerful regulatory mechanism in the transformation of dendritic Ca2+ signals into morphogenetic signals for the dendritic cytoarchitecture of principal neurons under pathophysiological and probably also under physiological conditions.How could this singularity with respect to other Caeraction . For ins(InsP3R) –60. The  release ,60. At tchannels ,62. This6 cotransformants were screened, and 108 clones were picked, which turned blue within 6 h in the initial test and after retransformation. Eight of these clones were found to encode a novel protein. Interactions were scored for ß-galactosidase activity by a colony lift assay. Binding activity of different constructs after retransformation was evaluated in three independent experiments.Yeast two-hybrid screening was performed as described previously . LibraryA rat brain hippocampus cDNA Lambda ZAPII library (Stratagene) was screened with a cDNA probe encompassing the first 400 bp of Jacob's open reading frame. cDNA labelling, filter hybridization, and subcloning were done using standard procedures . Cloning1–230 and GST-J253–404 were used to immunize two rabbits and one guinea pig each. Specificity of the antibodies was tested on immunoblots of crude rat brain homogenate by preabsorption of the antibodies with corresponding N-terminal or C-terminal (J262–532) MBP fusion proteins or with affinity-purified antiserum.Two peptides (aa 285–299 and aa 300–314), the GST fusion proteins GST-JImmunohistochemistry and immunocytochemistry were performed essentially as described previously   linked by a 9- or 10-bp–long stemloop sequence were obtained from Biomers. Sequences were as follows: nuclear Jacob knockdown (RNAi-NLS: 5′ AGA ATG ATT CCG CGT CTG TAA 3′/bp 892–912 of the Jacob cDNA); nuclear Jacob scrambled control (scrRNA: 5′ AGA TAT AGT CGC CGT CTG TAA 3′); all Jacob isoforms' knockdown (PAN-Jacob: 5′ TGC TAC TAG TTA CAG TGT AGA 3′/bp 390–410 of the Jacob cDNA); all Jacob isoforms' scrambled control (5′ TGA TAG GTC TAT ACG AGT TCA 3′), Caldendrin sRNAi ; and Caldendrin scrambled (5′ AGA ATC CTA AGA CAA GTG CAG 3′). Forward and reverse oligos were annealed, phosphorylated, and cloned BamHI, HindIII into the pRNAT-H1.1/Neo vector (Genscript) for plasmid-based RNAi knockdown. COS-7 cells were cotransfected with Flag-Jacob or Flag-Jacob-Myc/His and the RNAi expression vector. Cells were harvested 2 d after transfection and the samples solubilised for SDS-PAGE.3 plates to 90% confluence and cotransfected with the shRNA-FUGW H1(+) (10 μg), the VSVg (5 μg), and Δ8.9 (7.5 μg) vectors using Lipofectamine 2000 according to the manufacturer to produce competent virus particles. For virus production, cells were grown in Neurobasal medium supplemented with GlutaMAX and B27 at 32 °C and 5% CO2 overnight; the medium was changed and virus harvested 48–60 h after transfection. Sterile-filtered virus was directly added to primary cortical neurons at DIV0. After 3 wk, cells were either fixed with PFA for immunostaining or harvested and prepared for SDS-PAGE.For lentiviral transfections, double-stranded, phosphorylated oligos were cloned BamHI/BglII, HindIII into the pZ-off vector and further subcloned EcoRI, AccI/BstBI into the FUGW H1(+) vector. HEK-293T cells were grown on polyD lysine-coated 10-cmpSFV-ΔMyr-Jacob-EGFP, or pSFV-EGFP after in vitro transcription were cotransfected into CHO-K1 cells with Lipofectamine2000 (Invitrogen) according to the supplier's manual. After 24 and 48 h, the culture medium containing the budded particles was harvested. Viral particles were concentrated by ultracentrifugation through 10% sucrose, the pellet was resolved in Tris-buffered solution overnight at 4 °C. Aliquots of the particles were stored at −80 °C after shock freezing. For infection of primary cortical neurons, the particles were activated by chymotrypsin and further diluted with OptiMEM. High-density cortical cultures were infected at DIV16 and harvested 24 h later. Neurons were homogenized in 20 mM Tris buffer containing protease and phosphatase inhibitors, and solubilised in SDS buffer. The protein concentration was determined by amido black test, and equal amounts were loaded for SDS-PAGE.For the preparation of Semliki Forest particles and infection of primary cortical neurons, pSFV-Helper2, p < 0.05 was considered statistically significant.Statistical analysis was performed with ANOVA and subsequent Bonferroni's Multiple Comparison test. Data are presented as mean ± standard error of the mean (SEM). A level of Details about the computer models can be found under Supplementary Materials and Methods in Figure S1(1.5 MB TIF)Click here for additional data file.Figure S2(2.2 MB TIF)Click here for additional data file.Figure S3(898 KB TIF)Click here for additional data file.Figure S4(360 KB TIF)Click here for additional data file.Figure S5(570 KB TIF)Click here for additional data file.Text S1(73 KB DOC)Click here for additional data file.http://www.pdb.org/pdb/home/home.do) accession number for the structural model discussed in this paper is 1wdc.The Protein Data Bank (


Molecular imaging is a newly emerging and rapidly developing biomedical imaging field in which the modern tools are being married to depict noninvasive in vivo cellular and molecular processes sensitively and specifically, such as monitoring multiple molecular events, cell trafficking and targeting. The goals of this field are to develop technologies and instruments for studying biological and medical processes as well as diagnosing and managing diseases better. Although rapid progress in the fundamentals and applications make molecular imaging become an important tool for biomedical research in recent years, many difficult problems and challenges remain. Discussing the problems and challenges in detail and illustrating recent progress and future direction, the special issue collects the high-quality, peer-reviewed, original research papers in the area of molecular imaging.Novel molecular imaging theories and algorithms, new molecular probes, multimodality molecular imaging prototype systems and experiments, and final clinical applications are introduced mainly in this special issue. In molecular imaging theories and algorithms, genetic algorithm-based optimization tool is used to improve the accuracy of the diffusion model in strongly absorbing media by adjusting the optical parameters. Furthermore, a penalized linear and nonlinear combined conjugate gradient method, a fast preiteration algorithm based on the generalized inverse matrix, and a Monte-Carlo-based network method are also proposed for light source reconstruction in optical tomography. Considering the importance of molecular probes, synthesis and bioconjugation of gold nanoparticles as potential molecular probes for light-based imaging techniques are described in this special issue, and the nitroimidazole-based thioflavin-T derivatives as cerebral ischemia markers are evaluated in vivo. In order to test the feasibility and effectiveness of imaging theories, algorithms and probes, molecular imaging prototype systems should be designed, constructed and employed for small animal or phantom imaging. Thus, multimodality fusion near-infrared optical tomography Systems with highly sensitive CCD camera and photomultiplier tube can be consulted respectively in this special issue. Furthermore, a novel confocal optical system design and a dual laser confocal scanner have been developed for molecular imaging applications of biochip. In clinical applications, the advantage of PET and CT integration in examination of lung tumors is analyzed. Moreover, several innovative processing methods of molecular image are also presented in this special issue. In conclusion, this special issue covers recent important advances in molecular imaging field.


Madras in the eighteenth century was a site of continuous warfare sparked mostly by trading interests. This paper studies how these influences of hostility and commerce shaped the medical establishment of the English East India Company. It begins by analyzing the struggle of the medical establishment to cope with military and logistical requirements; it then shows how the Coromandel trade provided a peculiar dynamic to the practice of medicine in Madras. By aligning the history of medicine with that of trade, the paper traces the parallel trajectories of intellectual and material wealth. The development of modern medicine is seen as a process of adjusting to and engaging with diverse ideas and items—sometimes co-opting them, sometimes realigning them in new modes of production. The port of Madras (Chennai) on the Coromandel Coast of India was founded in 1639, and Fort St. George in the 1640s, by the English East India Company , a joint-stock corporation of English merchants, as part of their network of warehouses or “factories” protected by military settlements and forts across South and East Asia. ThroughoutHistorians of the emergence of medical institutions and hospitals, particularly those of the EEIC, from the late seventeenth century have mainly focused on the theme of war, sometimes characterizing the medical practices as “military medicine.”In the first part of the paper I discuss how the medical establishment was created by the nature of the EEIC's settlement and military activities in Madras. The next section demonstrates how commerce, the chief concern of the EEIC, was also crucial to the growth of that establishment. The practice of medicine in Madras needs to be mapped along the trade routes, in the items of barter and in the markets of exchange, to construct a history of treatment enriched by trade.Finally, I will show that medicine in Madras has to be understood in terms of the shifting political, military, economic, and intellectual dynamics on the Coromandel Coast. At a more general level I will show how modern medicine developed as an attempt to adjust to and engage with the multifarious collections of ideas and items—sometimes co-opting them, sometimes realigning them in new modes of production. MadrasC. A. Bayly has suggested that the financial and military momentum of the EEIC's army was the mainspring of British expansion, particularly in India, during the early eighteenth century.An early concern of the EEIC was to ensure the health and discipline of its European soldiers and seamen at its foreign settlements. In 1664 William Gyford and Jeremy Sambrooke wrote to the governor of Madras, Sir Edward Winter, saying that a hospital and regimented health care were necessary to treat the English soldiers:soe the fresh souldiers which came forth this year takeing up their habitation in the bleake winde in the hall, fell sick. Four of them are dead: about tenn remaine at this time very sick and complaine (and it seems not without Reason) that the wages are not suffcient to supply them with the necessary now in this time of their sickness. Soe, Rather than see Englishmen dropp away like doggs in that Manner for want of Christian Charity towards them, we have thought it very Convenient that they might have an house on purpose for them, and people appointed to looke after them and to see that nothing comes in to them, neither of meate nor drinke, but what the Doctor alloweth.Thus the earliest hospital in Fort St. George was established on 16 November 1664 with John Clarke as the first surgeon. In 1688, for want of space, it was moved to the northern end of the barracks.dubashes,As pointed out by Mark Harrison, it was only in the mid-eighteenth century that the expansionist policy and “open warfare” of the Company led to the employment of regular forces in Bengal and Madras Presidencies.cooliesdubashes were to make and prepare dressings as well as to help in preparing medicines by pounding the mortars. Local women were to clean the wards and provide the hospital with water, and a “head coolie” was to supervise the Indian subordinates.Subsequent wars also created the need for enlargement of the hospital facilities. In November 1758 the surgeons Robert Turing, James Wilson, and T. S. Hancock sent a comprehensive proposal to George Pigot, president and governor in council of Fort St. George, for a new hospital—theThe Military Board initially approved most of the submissions, but war caused these plans to be shelved. In May 1771 the Board finally resolved to accept the proposals for the construction of a new hospital building, following closely the administrative and architectural outlines of the 1758 plan.The practice of medicine in Madras also had to engage with battlefeld and logistic concerns. Victory did not ensure well-being in a hostile land. In January 1760 the English recorded one of their most decisive victories in Deccan, at Wandiwash against General Lally's French troops. The victorious General Eyre Coote, however, found the battle scenes “dreadful” due to the lack of medical support in the field:Such a multitude of poor objects, and not in my power to give them the least assistance for want of every one necessary requisite for an hospital. I make no doubt upon this representation you will do everything humanity can direct. If it is possible to send surgeons and proper people from Madras to attend the wounded here who are very numerous, you may by that means save theThe lack of either adequate flying field hospitals or mobile staff in India made the transportation of medicines from the hospitals to the battlefields, and of the sick or wounded from the battlefelds to the hospitals, a major problem. The situation was made worse by the favorite tactic of both Hyder Ali and Tipu Sultan of starving out the British armies by cutting off their supplies.doolies:doolies the sick had to walk to the hospital, a distance of two to three miles, under the hot sun; this problem of logistics was also related to the confusions regarding command and authority faced by English surgeons in Madras, for he added: “I also request to know if these Dooley boys are to be under my immediate direction, the difficulty of getting them when wanted, having often occurred from the Dooley boys believing that they are to obey no orders given them but from the person who pays them.”doolies and bullock-drawn carts to carry the sick and the wounded.Senior Surgeon Colley Lucas complained about the lack of coolies to carry the sick in the field.Nightingale with the annual supplies of medicine for Madras was lost at sea, crisis struck the hospital: the president wrote to the Board that the surgeons had frequently told him “of the want of Medicines in the Hospital occasioned by the loss of the Nightingale,… It is therefore Agreed that the same be purchased allowing twenty five per cent advance and that the amount thereof be paid for out of Cash.”The supply of medicines was the main concern in the field and the provincial hospitals. The main provision for the Madras hospital came in the form of medicine chests from Europe; these were often destroyed or lost on the long voyages from Europe, and the surgeons then had to re-Complaints were also made about the poor packaging of medicines from Europe. In 1774 Gilbert Pasley and James Anderson wrote to the Board about the European medicines received for 1773, which had arrived “much damaged on account of the careless and injudicious Package of them,” and they added that the “supply of surgical Instruments is likewise greatly deficient.”The Hospital Board beg leave to take this opportunity of informing Government of the imperfection and indeed the inutility of several of the Medicines brought from Europe, especially Cantharides or blistering flies, the efficacy of which is so considerable in many critical disorders, and the consequent disappointment so great.The situation was particularly grave in the provinces of the Presidency, which depended on Madras for their supply of European medicine. In 1787 John Duncan, assistant surgeon with the Battalion of Native Infantry, represented the “great want” of a medicine chest to carry medicine easily and to protect it from pests.But the “field” of medical practice and treatment in the Madras Presidency stretched beyond the zones of military confict and bloodshed, often to the “Country” (in its Anglo-Indian sense), which was also the source of supply for medicines. This is where the story of medicine at Madras is linked to the main concern of the EEIC, its trade in the Indian Ocean world.The practice of medicine beyond Europe in the eighteenth century posed many challenges for the Europeans. Apart from war, the climate and the customs of the Coromandel Coast also troubled the European constitution and conscience. Charles Curtis, a surgeon from England who briefy visited the hospital during 1782–83, found it and its surroundings rather inhospitable:toady, was all we could procure for our scorbutics. Another source of unhealthiness arises from the situation of the town, and fort of Madras itself, built upon a low, flat and sandy beach, and surrounded almost entirely by stagnant swamps, or ponds of stagnant water, which are not only a receptacle for every sort of corruption and filth, but have their edges, as well as the beach on the other side, covered at all hours of the day with the natives, in the act of relieving nature from their burdens, to take advantage of the water for washing afterwards, which is their invariable custom. What a source of putrid exhalations under a vertical sun!The country round was more than once overrun, and plundered by the horse troops of Hyder Ally. So that our supply, even of fresh meat, and much more of vegetables and fruits, was by far too scanty. And a few imperfectly cultivated and unripened fruits, with the cocoa-nut juice called Transactions of the Royal Society.But where there was grime, there was also bounty. Throughout the years of war the EEIC was establishing itself as the principal trading power in these parts. Madras, as its stronghold in this expansion, was the hub of various networks and arbitrations and, as suggested by some historians, a site not of degeneration but of “vitality.”The EEIC itself was deeply entrenched in the social life of Madras and its surroundings in the early decades of the eighteenth century. The local nayak or the nawab of Carnatic had little visible control over Madras, a port city established by the British. As Holden Furber described it: “Fort St. George was much more impressive than Fort William [at Calcutta]. The Governor and Council actually ruled over a sizable local population, composing caste disputes and making authority felt well outside the so-called ‘white’ and ‘black’ towns of Madras, especially in the countryside where the weavers of the ‘Company's cloths’ lived.”While problems of infrastructure and logistics plagued the medical establishment of Madras, surgeons from very early on had discovered other sources for the supply of medicines. The eighteenth-century history of the hospital shows several instances where surgeons depended on the local bazaars. By the middle of the century, Country medicines or bazaar medicines, as they were referred to, had become a regular part of the supply for the establishment and the surgeon general's records maintained separate requisitions for them. One of the main proposals of the 1758 report from the surgeons was to have separate rooms for European and Country medicines.dubash for his shop to assist in the preparation of medicines and to make up dsressings, The surgeons' conversance with local supplies is evident in the 1762 proposal of the surgeon James Wilson to personally supply the hospital with cots, clothing, Country medicines, servants, and all other necessities except “Europe Medicine”; he also proposed to hire a Private procurement, however, continued. In 1796 the surgeon Bernard McMahon wrote from Ceylon that he had bought medicines from the bazaar to treat his patients suffering from ulcerous putrefaction.One reason for the growing need for bazaar medicines was the increasing numbers of Indians recruited in the forces, who were generally treated with indigenous medicines. During 1765 a major expansion of the Native Infantry took place, and at about the same time the government sanctioned the appointment of a “Native” doctor to each sepoy battalion with the pay of a havildar.What constituted this “Country medicine”? At one level, it can be defined in terms of supply: anything that was procured locally and was not sent from Europe constituted “Country medicine.” Behind “Country medicine” were the local “native” practitioners and informants on whose“Country trade” was the term used by the Europeans for the local trade of South and Southeast Asia in which they participated increasingly, often privately, throughout the eighteenth century. One of the important successes of the European trading powers was their ability to divert this trade not only to the ports controlled by them but also to the goods of their interest.Philosophical Transactions, James Petiver referred to an earlier discovery of a root in the East which was found to be particularly useful in epileptic and convulsive diseases, justifying, according to Petiver, the need to “better understand” such medicines and “to have them in greater plenty, so that even the poorer sort of People may receive beneft by it at a much moderate price, and the Merchant receive advantage by trading in an new Commodity.”During the eighteenth century the history of the hospital in Madras was shaped as much by European trade to and from the Coromandel Coast as by the requirements of the Carnatic and Mysore wars. In the early decades of the century doctors like Robert Turing were generally engaged in private trade.Swietenia febrifugaScience and trade were thus closely interlinked. The main Madras contact of William Roxburgh, an EEIC botanist posted at Samulcottah  in Andhra Pradesh, two hundred miles north of Madras, was Andrew Ross, the most prominent trading agent in that city involved in country trade. Ross procured books in Madras to send them to Samulcottah for Roxburgh; Roxburgh sent Ross a breadfruit plant, to be local medicines procured and used in South Asia and the West Indies . My suggestion here is not simply that Country trade amplifed the use of Country medicine. Rather, this comparison can be used as an analytical tool to understand how trade can be seen as a metaphor for medicine, particularly for this period; how it signifes European involvement in the “native” or the local; and how that shaped the identities and content of both. In the Oxford English Dictionary, the “Anglo-Indian” connotation of “Country” is defned as “Of or belonging to India (or other foreign country), as distinguished from European; native.”New Account of the East Indies (1727) on trade in the Indian Ocean region.But there were also other links, less obvious, between “Country trade” and “Country medicine.” The term “Country medicine” was used by Europeans for the Essays, Medical and Experimental.The history of eighteenth-century Country trade is rather checkered. Even before the Europeans established themselves in these parts, thereOn the Coromandel Coast and in Bengal, Country trade became prevalent in the eighteenth century, increasingly mingled with the trade with Europe. According to Furber, it was the commercial revolution of the early decades of eighteenth century that stimulated the growth of European participation;Materia Medica of Hindoostan published in 1813 by Whitelaw Ainslie  describes the complex lineages of bazaar medicines, which were “either the produce of Hindoostan, or…brought to it from Asiatic countries, and are to be met with in the Bazars of populous Towns; including many Drugs of the Tamool, Arabian, and Persian Materia Medica.”The medicines that the British surgeons found in the markets of South India did not come only from the neighboring forests: Country medicine was sometimes “Country trade” medicine as well. The products had a long and syncretic history of trade and network, often being brought over from other regions and used locally as medicinal ingredients. The Europeans similarly used them as ingredients for their own medicines, and also bought “pills,” barks, and roots commonly in use in the surrounding regions. The Important sites of Country trade like Machilipatnam also became signifcant points of procuring Country medicine. In 1788 the surgeon George Binney wrote from Machilipatnam to James Anderson, mentioning that “Ginglee (Sesame) Oil” used in the composition of ointments could be found there, for sesame grew widely and the oil could be procured locally in a state fresher than that from the Presidency.I believe I might venture to say almost every article of country medicines is to be purchased in the Bazar here, and I may add at almost every station of my department north of the Kistna, of as good a quality, at as cheap a rate, and some of them cheaper and fresher than at Madras. … it seems an expedient highly necessary to be adopted; and will be calculated to do credit to practice, by securing the very important end—of everything being of the purest kind, & of the best quality.Manual of Materia Medica because the Medical Board wanted to ascertain whether supplies from Europe could be replaced by “articles indigenous in the country, or by cultivating exotics in the most suitable climates of the plains and mountains of North India.”The government, too, had realized the need for local procurement of medicines. In 1789 the Court of Directors wrote from London that itAcacia senegal and A. seyal, leguminous trees from Egypt),Plantago ovata), aloes , asafoetida ,Glycyrrhiza, mostly from the East, but also found in Spain and Italy), cinnamon, China-root ,Caryophyllusaromatics, from the Moluccas),Camphora officinarum, a tree indigenous to Java, Sumatra, Japan, etc., and from other lauraceous trees, brought to India from China by Country trade ships),Daemomorops draco), fennel seeds, gamboge ,Sesamum indicum, from India),Styrax benzoin, a tree of Sumatra and Java),Alpinia and Kæmpferia, introduced to early medieval Europe by the Arabs),Commiphora, in Abyssinia),The substances listed as “Bazaar Medicine” in the hospital records are recognizable as items of trade as much as of treatment, and they suggestWhitelaw Ainslie detailed the Tamil physicians' preparations of nitrous, muriatic, and vitriolic acids for medicinal purposes.We are informed by Captain Turner, that, at Tessoolumbo in Thibet, Cinnabar is found which contains much Quicksilver: and I perceive by the little volume entitled “Remarks on the Husbandry and internal commerce of Bengal” that Mercury thus mineralized might be considered as one of the export articles of trade from Hindoostan; the greatest part of that valuable metal, however, which is exposed for sale in these provinces, is brought to us from China; where it is procured, both in its native purity (See Abbe Rochan's “Voyage to the Madagascar and the East Indies” p. 365–66) and combined with Sulphur.Thus while the bazaars were being seen as alternative sources of supply for medicines, European participation in trade and therapeutics had further syncretized these traditions. And Madras, the “open roadstead,”In July 1788 Mr. Strange, a surgeon near Tanjavur, wrote to Governor Archibald Campbell in Madras about a “native” practitioner, introduced to him by the Danish missionary Frederick Schwartz, who could prepare some kind of “snake pills” that apparently cured people bitten by snakes. Both Strange and Schwartz wanted this man and his pills to be examined by doctors in Madras. The governor asked for the man to be sent to Madras, where the Hospital Board was to make trials of the pills “for the purpose of having the merits of his discovery ascertained.”Analysis led to intervention. In September, William Duffn and other surgeons wrote to the Hospital Board that although the results of repeated trials were successful, some of the components raised questions, and they could “only recommend the Government to leave every practitioner to administer remedies, as his own judgement may direct as heretofore in cases of venomous bites.”A simple adoption of these pills, which had promised a major cure, was not seen by the European medical establishment in Madras as an option, and analysis of the ingredients became vital to this European engagement with the Tanjore pills. In October 1788, after preliminary analysis, the Military Board approved and encouraged the use of the pills in cases of snake and mad dog bites.that you will without loss of time draw up an accurate account of the names and proportions of Articles composing these Pills specifying in a particular manner the Malabar and English names of each Article, as well as the botanical names of such plants or Roots as are used in the Composition of these Pills and the manner of using the Pills. …The Board at the same time direct, that you will publish the same without delay in the Madras Courier, and that a quantity of these Pills may be immediately made up and sent to every Station under the Presidency.Account of Indian Serpents Collected on the Coast of Coromandel published in 1796 from England he described his experiments with these “Tanjore Pills” as inconclusive, and the pills were found to be rather ineffective.In November, James Anderson in his minute on the pills produced a detailed list of their ingredients and added that it was indeed arsenic that put their usage in doubt. He asked for a ban on the internal use of the pills, “wishing as I do to expel for the sake of humanity such noxious Drugs from our Materia Medica.”The blend of medicine and ideas in Madras was indeed intriguing. Thus sidelined in Europe, the pills and their ingredients had an interesting impact in India. In 1789 William Jones wrote to Anderson at Fort St. George, who seemed to have meanwhile changed his mind, about his own successes with arsenic:I was much interested in your papers concerning the Hindu pills for the cure of persons bit by venomous snakes. Till I saw Duncan's Medical Commentaries, I had the same abhorrence with yourself to the use of arsenick in medicine, and had resolved to suppress a paper of a Native Physician concerning the cure of the Elephantiasis by small quantities of arsenick; but when I found that it had been safely given in England for intermittents, I ventured to print it in the second volume of our Transaction: … Poisons must, no doubt, be administered with extreme caution; but if we banish them from practice, what will becomeBut Russell's judgment largely put an end to the debate on the pills. Thus while spaces for new ideas and ingredients were being created in the peripheries, confrmation from the metropolis remained important. This duality becomes evident in Anderson's reply to Jones. Urging caution in the use of arsenic or any other poison in medicine, Anderson added:“[that] the natives of this country are better acquainted with the use of Simples than any of the Peoples may be readily admitted by all who consider the situation the stimulation of the country and the nature of their food and there is great probability that Chymistry has flourished here.”The “Tanjore pills” seem to have retained their ambiguous position for almost another century. In 1873, T. Lauder Brunton and J. Fayrer wrote that although they enjoyed a “large amount of popular confidence,” “when brought to the test of carefully conducted experiment, [they] failed, as might have been expected, to give any favourable result.”It is worthwhile here to go back to Duffn's earlier defense of the pills and arsenic:[sic] demonstration have in many cases proved it may be given with safety, besides the internal use of arsenic is by no meansas we are still ignorant of the analysis of the exotic Plants, and other vegetable matter which are compounded with those minerals and how far they may counteract the Violence of the Arsenic we should not hastily condemn a Remedy where experience & occular Materia Indica with the common European reservation about the chemical skills of the local practitioners: “The preparations of mercury found in use amongst the Tamool practitioners give us but a poor opinion of their knowledge of chemistry”; but he then added: “Yet, after all, however much we may be inclined to smile at some of their strange mixtures, it must be confessed that the characterizing principles are generally correct, and that, every thing considered, there is, in the present state of knowledge amongst the Vytians and Hakeems,not always intentionally, to induce by its use most frightful salivations.”The conflict between the ocular and the analytical that is evident in Duffn's words in fact underlines a larger tension, both between different modes of scientifc credibility and between the metropolis and the periphery. This was manifested in the duality in the work of European scientists in India: the Madras hospital and its surgeons had continued to depend on the local bazaars and practitioners in this era of a distinct medical research and production in Europe. Ainslie began his the medical works of the Hindoos are neither to be regarded as miraculous productions of wisdom, nor as repositories of nonsense. Their practical principles, as far as I can judge, are very similar to our own; and even their theories may be reconciled with ours, if we make allowance for their ignorance of anatomy, and the imperfection of their physiological speculation.This is an important problematic of these interactions: they were as much to treat the sick in Madras as they were advertisements of peripheral research and novelties for Europe. The duality in the works of Ainslie,Agashtier Vytia Anyouroo. John Forbes Royle, born in Kanpur (in north India), entered the service of the EEIC as assistant surgeon and “made collection of all the drugs procurable in Indian bazaars, tracing them as much as possible to the plants, animals, and countries, whence they were derived”;Edinburgh New DispensatoryAnnals of Medicine.The link between the local and the metropolitan is situated here. Signifcantly, Ainslie categorized his list of bazaar medicines as a part of “British Materia Medica” that “could be procured in the Bazars of Hindoostan,” and located them as much in the local markets as in the texts of Tamil scholars like Agashtier, These promotions and publications were informed by a search for legitimacy and autonomy by surgeons and physicians throughout the empire.experience a local knowledge of the diseases of this climate.”In January 1775, Pasley and Anderson made an appeal that young medical assistants sent from England should possess certified qualifcations.“peculiar to the Coast of Coromandel,” a “medicine of greater power” was needed.Pasley had played an important role in asserting the autonomy of medical research at the Madras hospital. Part of this independence was sought from the physicians at home. In 1771, when he was recommended by a London physician advising the Court of Directors to use castor oil in treating infammation of the liver, Pasley replied: “I am sorry to observe it as a proof of how ready Men of reputed eminence in Physic are to precipitate on the Work an opinion regarding the Virtues of particular Medicines, without a suffcient personal Experience of their effects; consequently the only conclusive Test, of the certainty of their powers and operations.”Interestingly, Gahagan, who later joined the establishment, becoming first the chief surgeon at Vellore and then a member of the Medical Board, asserted the same sense of the novelty of medicine in Madras. In 1802, another centralization and militarization of the services was being undertaken. Gahagan, as the Second Member of the Medical Board, produced a note of dissent to a proposal by the Court of Directors to dispose of the General Hospitals of the Presidency in order to curtail costs. His primary defense of the hospitals was that they provided ideal training grounds for British surgeons in this country:they open an ample field for young practitioners upon their arrival from Europe in this Country; They afford them an opportunity of acquiring a knowledge of local diseases and the best mode of treatment adopted in their care under the long experienced practitioners whence they may proceed better enabled to act of themselves, with confidence of success whenever their services may be required in detached situations.To Gahagan, Madras was the site where theory was put to test by experience, “for although a young man may come out well grounded in a theory, it still requires practice, experience and diligent observation to getGahagan's final point was that the hospitals were also a training ground for indigenous practitioners: “General Hospitals are the most useful and preparatory Asylums for Native Medical Servants, such as Assistant Apothecaries, Dressers, nursing attendants, and for various other purposes.”This highlights the other aspect of the search for distinction in the city of Madras: that from the region itself. The emergence of Madras as the principal site of medical innovation in the region was hardly a coincidence: it was the British stronghold on the coast. This particular metropolis/province confguration in scientific expertise was linked to commercial enrichment and displacement, a topic with a complex historiography. The relationship between metropolitan and provincial science in Europe has been well documented.documented.But what remains outside the scope of this historiography is the analysisInterestingly, here too networks have been traced: those connecting agrarian regions with the world economy in the centuries before 1800.What is obvious from a survey of the historiography of the eighteenth century is not a picture of a lack of dynamics or “vitality,” but one of shifting dynamics. Subrahmanyam captures the contours of this change.Thus the historiography of the commodifcation of curiosities, of trade and treatment, of commerce and science, and of material culture and knowledge, as developed for science in Europe, needs to be placed alongside another historiography of material culture: that describing the economic and political histories of shifting dynamics and sovereignties in areas beyond Europe. A link between the histories and historiographies of displacement and accretion thus needs to be forged. In the next section I explore these possibilities.trust, which were shaped by the link between virtue and expertise.virtue and expertise.By sending the Tanjavur physician along with his pills to Madras, Schwartz had confrmed that the Madras hospital by the 1780s, like the city in trade, had become the center of medicine along the Coromandel Coast. Similarly, Benjamin Heyne of the Tranquebar mission, a keen natural history enthusiast, had also migrated to the Madras establishment to pursue his scientifc projects.Much of the centralization process in medical practice was prompted by the need of the military establishment to organize the medical establishment around Madras. In April 1760, for better integration among several hospitals in other parts of the Presidency, the Military Board resolved that one “General Hospital” was to be maintained in Madras, and the sick and the wounded at the garrisons at Arcot and Chinglepet were to be sent to Madras.In April 1786 the Medical Department and the Hospital Board were formed, with James Anderson as the physician general.Madras being the Capital of all the Hon'ble Company's possessions on the Coast, & where all persons coming from Europe, to serve under this Presidency, disembark,… it appears necessary to establish a General Hospital at this place for their immediate reception, in which the sick may receive every assistance that can be derived from the experience of the surgeons who from their long residence in this Country, & their knowledge of its productions, are best acquainted with the proper remedie to be used for the recovery of the Sick and also where such remedies are to be procured without delay.Militarization also implied the curbing of the role of the medical establishment. By the General Order dated 14 April 1802 the Medical Board was removed from nonmedical activities such as administration, including the right to reward or recommend someone for promotion or remuneration, and was to restrict itself to “those objects of Medical Practice, which belong to its immediate province.”The medical establishment was opposed to such a military takeover. This is where the otherwise correlated processes of centralization and the search for autonomy came into confict. In response to these orders the Hospital Board expressed its “dismay” and “concern”: it cited the example of England, where the Medical Board recommended the appointments of medical staff, and it argued that the Medical Board had played a central role in running the entire medical establishment of the Presidency, a role that the Court of Directors had previously recognized.With medical men therefore detached and living entirely in military Societies, cut off from the Head of their Department, and deprived of all hopes of success in the Service, by any attempts to recommend themselves, in the only proper way for Medical Men to do it is not difficult to foresee how much by such a system, the relish for professional pursuits in many young Practitioner must be lost, greatly to the future falling back on the Hon'ble Company's Medical Establishment and consequently to the prejudice and distress of the pubic service.Some boundaries, however, were put up by the surgeons themselves. The development of the Madras hospital as an important training site for indigenous practitioners was also a process in defining therapeutics. There were simultaneous efforts toward reducing the dependence on local practitioners, which sometimes refected an emerging distrust of them. This had interesting connotations in terms of both knowledge and politics.On the one hand, this was the period of Europe's assertion of its superior scientifc achievement vis-à-vis other regions, engendered in the duality discussed above. On the other hand, the Company was now establishing itself as the dominant regional power and reducing its dependence on local informants. This brings into question how far we can understand the interactions between the Europeans and Indians as “dialogues” and “conversations.” Scholars like Eugene Irschik, Thomas Trautmann, and Philip Wagoner, describing the nature of communication between the colonizer and the colonized in eighteenth- and nineteenth-century South India, have sought to assign a more “active” role to the colonized, to show that the “indigenous intellectuals in reality contributed actively to the process, and that colonial knowledge was thus produced through a complex form of collaboration between colonizers and colonized.”The same year (1792) that the Hospital Board appointed Indian contractors for the supply of bazaar medicines, it also ordered that Indian “servants” (assistants or “black doctors”) were not to be allowed to prepare medicines or dress the wounded soldiers, for they were considered incompetent in these areas.The government, more than the medical establishment, had become suspicious of local assistants—even those trained by Europeans. In 1800, when the Medical Board wrote to the government about appointing a “Malabar Assistant” who had been “brought up” by a French chemistdubashi institution, “typified in the European enclave … [as] a creation of the eighteenth century,” is an important instance of this changing scenario in Madras.dubashes functioned as guides, advisors, brokers, and even moneylenders to the EEIC offcials. Their preeminence in the mid-eighteenth century over other local merchant groups indeed symbolized the ascendancy of Brahmin and Velalladubashi connections.dubashi infuence had almost withered away and the term disappeared from the official nomenclature.need to dissociate themselves from these groups.ability of the EEIC to replace them: prohibitions regarding private trade reduced dependency on dubashes as agents, and linguistic training provided to junior civil servants in Madras reduced the need for translators or intermediaries.The There were also limits to the surgeons' search for eminence and autonomy in medical research in Madras. To begin with, the status of the surgeons within the EEIC hierarchy was rather low. Financial rewards forif their heads there are turned, give us due Notice, that we may call them home again and supply their Places with Men of more humble minds, though perhaps not inferior Talents.You tell us that the Salaries of our Surgeons must be enlarged if we expect or desire to have Men of Ability in their profession; The Surgeons that we send abroad to our Capital Settlements are always acquainted with their Salaries and Emoluments, and we find no difficulties in having able Men of that profession, as well as all other Branches of our Service, There was little infrastructure for the large-scale production of “Country” medicine to support the needs of the increasing army. Pasley responded to the wish of the Court of Directors that medicines be prepared in India by showing that such a plan was impractical due to the want of utensils, chemical laboratories, and trained assistants.The history of the growing prosperity of the Society of Apothecaries in London is related to these developments in Madras. In 1766 the Society obtained the monopoly for supplying medicine to the EEIC. This became the core of its business, and a period of flourishing profits in the late eighteenth century followed. In the 1820s it was the chief supplier of the huge Indian army of approximately 300,000 men. The Society boasted: “the whole army of India (we believe exclusively) is served under the direction of the Hon United India Company from our establishment.”but may ultimately increase the demand for European Medicines to an object of commercial importance.” One incident serves to illustrate the nature of medical practice in Madras at the end of the century. In 1800 Thomas Evans, who had very recently joined the Madras establishment as an assistant surgeon, made a proposal for the establishment of a dispensary in the Black Town. Evans had two justifications: first, such a dispensary would help to promote European medicine in these parts, thereby making commercial proft for the EEIC; second, it would help to suppress all “irregular practitioners and vendors of medicine in Black Town.”Remarks on the Uses of Some of the Bazaar Medicines and Common Medical Plants of India, written from England, expressed the need to “introduce” bazaar medicine to these people in the colony: to the missionaries working there, the resident Europeans and Anglo-Indians, and the Indians, “the daily increasing array of educated Natives, who as a class, readily avail themselves of every scrap of knowledge drawn from trustworthy European sources, which tends to throw light on the products and resources of their native land.”By the middle of the nineteenth century, bazaar medicine had come a long way in its metropolitan participation. Edward John Waring , in his celebrated Eighteenth-century Madras was a site where both “Western” and “Eastern” medicines were subjected to trial. War, isolation, cultural exchanges, and climate had expanded and redefined the possibilities of these practices. On the other hand, networks, collaborations, and negotiations were vital to European endurance in these distant lands, and trade was both the means and the metaphor of these exchanges. The “rise” of the bazaar from the eighteenth century has been seen as a new “distinct world” of sophisticated negotiation and bargaining developed by the indigenousSince it was not just medicine that was being bartered here, the general economic history of Coromandel trade becomes relevant. A comparative analysis of Britain and South India in the eighteenth century in terms of wages, prices of grain, and agricultural productivity shows that the image of “impoverished masses” of India, as painted by European travelers, was misleading. Such an analysis has not only challenged concepts of Oriental despotism and static and self-sufficient villages, but also has hinted at the need to rethink Europe's industrial revolution, which could be seen as a response to the “economic dynamism outside,” Britain's answer to India's competitive challenge.On the other hand, as one of the largest employers in eighteenth-century Britain, the EEIC shaped British middle-class earning and spending. The EEIC clerks were more able to earn higher salaries, compared with other banking, commercial, and governmental employees between 1750 and 1850, and the economy generally benefted from vigorously escalating salaries in these classic years of industrial revolution, which in turn led to domestic demand for the goods of the new industrial sector.dubashes, “black doctors,” surgeons, and arsenics, its shifting dynamics and sovereignties, and its tributes of dried seeds and texts sent to Europe—can thus inform us more about the researches and resources of Western science.It is necessary to align the history of medicine with economic history: not only to write an economic history of medicine, but also to align the trajectories of intellectual and material wealth. These triumphs of the eighteenth century contributed to the power of nineteenth-century Western science otherwise visible in its identifcation of microbes and vectors and in its eradicative and preventive successes in the tropics. This complex world of Madras—its


The association of the human herpesvirus-8/Kaposi's sarcoma (KS)-associated herpesvirus (HHV-8/KSHV) serology with various malignancies in Tanzania is not currently well established while previous studies were based on either PCR or immunofluorescence assays [IFA] but not with a sensitive enzyme-linked immunosorbent assay (ELISA). Selected archival diagnostic biopsies (n = 184) and sera from indigenous patients with KS (n = 120), non-KS tumors (n = 24) and non-neoplastic lesions (n = 40) at Muhimbili National Hospital (MNH), Tanzania, were evaluated by diagnostic histopathology, immunohistology [anti-HHV-8 latency-associated nuclear antigen (LANA)] and serology for HIV (ELISA) and HHV-8 (IFA and ELISA).About 66.3% (n = 122) cases including AIDS-associated Kaposi's sarcoma (AKS) (n = 93), reactive conditions (n = 28) and only one non-KS tumour were HIV positive. Endemic KS (EKS) patients were mostly males  who were less  predominant in AIDS-associated (AKS). A high (89%) percentage of patients with anti-HHV-8 antibodies was found in the cohort including the HIV positive (92%) cases, males (81.2%), KS patients (93%), non-KS tumors (92%), and reactive conditions (75%). All HHV-8 seronegative KS cases were nodular stage whereas both sera and corresponding biopsies from early stage KS were HHV-8+. Assay sensitivity, positive predictive value (PPV) and specificity were 98.6%, 93.5% and 16.7% for IFA and 93.5%, 98.6% and 50.0% for ELISA respectively.HHV-8 seroprevalence at MNH appears high as expected among AKS cases and males but also in non-KS patients. ELISA showed a combination of high HHV-8 sensitivity as well as higher PPV and specificity than IFA which however, showed higher sensitivity. The apparent stage-dependent, inverted serum HHV-8 immunoreactivity supports a notion of viral immune-segregation during KS development. Routine HHV-8 screening should be considered particularly in patients at risk of KS and for selection of blood/organ donations. The HIV and AIDS epidemic has dramatically increased the frequency of different malignancies particularly Kaposi's sarcoma (KS) and certain malignant lymphoma (ML) which are associated with the novel human herpesvirus type 8 (HHV-8)/Kaposi's sarcoma-associated herpes virus (KSHV) and have become a major health concern in sub-Saharan Africa including Tanzania -3. The pA total of 184 selected biopsies and corresponding sera available at MNH/MUHAS between 1990 and 2001 from indigenous African patients were included in the study. Of these patients, 120(65%) had KS, 24(13%) non-KS tumors and 40(22%) non-malignant (reactive) conditions. The male:female ratio was 2.75:1 for KS, 3:1 for non-KS tumors and almost 1:1 for reactive, non-malignant conditions Figure  and  were nodular stage Figure  with cut2 = 7.96, p = 0.0048)  and at the Immunopathology Lab . Furthermore, available biopsies and corresponding sera from non-KS tumors and reactive conditions from the same period were also evaluated. The biopsies of reactive conditions included all histologically non-malignant/inflammatory cases whose sera were available for analysis. None of the patients had received any antiretroviral (ARV) or anti-tumour therapy before the biopsy was taken.Routine hematoxylin and eosin (H & E) staining was done as previously described ,26 and mAvailable sera from patients with histologically diagnosed KS, non-KS malignancies as well as reactive conditions were comparatively evaluated for HIV and HHV-8 serology. Serology for HIV-1 evaluation (ELISA) was performed at Microbiology/Immunology MUHAS as previously described ,28.HHV-8 serology by IFA was performed [Swedish Institute for Infectious Diseases Control (SMI)] on cytospins of BCBL-1* cells  using pa2. To induce lytic gene transcription, cells were cultured with 20 ng/ml of 12-O-tetradecanoylphorbol-13-acetate (TPA)  as previously described, [® slides . The slides were fixed for 10 minutes in 4% paraformaldehyde (PFA) and washed before immunostaining as previously described [The HHV-8 infected body cavity-based lymphoma (BCBL-1) cells (kindly provided by G. Gaidano) were derived from a primary effusion lymphoma (PEL) ,31, and scribed, ,28,32 anescribed ,8.For HHV-8 antibody evaluation an IgG enzyme immunoassay (EIA) Kit  Biotrin International  was used. The Biotrin HHV-8 ELISA is a direct EIA based on the binding of HHV-8-specific antibodies to lytic peptide antigens coupled to microtitre test strips. Bound antibodies are detected by an anti-human IgG peroxidase conjugate and a 3,3',5,5'-tetramethylbenzidine (TMB) dark blue substrate reaction . The use of lytic peptide epitopes derived from various HHV-8 viral proteins ensures both a high sensitivity and specificity. The Kit has no detectable cross-reactivity with HIV/EBV antibodies. Control sera included those from known HHV-8+ KS patients while the HHV-8 infected BCBL-1 cells provided an internal positive control for IFA studies. The negative controls for both IFA and ELISA included sera from known HIV and HHV-8 negative non-KS patients and a buffer (PBS).Immunostaining with the avidin-biotin complex (ABC) immunoperoxidase technique was used on KS sections (5 μm) for detection of HHV-8 latency-associated nuclear antigen (LANA) to compliment histopathology and serology, as previously described ,26,33. NFor microscopy and microphotography an Olympus BX60, microscope with a digital camera (Sony DKC-5000) was used as previously described ,19,33.The EPI INFO 6 statistical software programme  was used. P-values of ≤0.05 were considered statistically significant. The Fisher exact test was used to test the significance level where the sample size was small.The authors declare that they have no competing interests.


Kawasaki Syndrome (KS) is an uncommon childhood disease with unknown etiology. It has been suggested that rotavirus infection may play a causative role in the development of KS.To examine potential temporal associations between KS and rotavirus infection, seasonal patterns of KS- and rotavirus-associated hospitalizations among children in California and New York during 2000-2005 were compared.Rotavirus hospital admissions were markedly winter seasonal, with very few summer hospitalizations. KS hospitalizations occurred year-round but also peaked slightly during winter and spring.The strong winter seasonal pattern of rotavirus clearly differed from the year-round pattern of KS hospitalizations. While the present study cannot completely rule out rotavirus as having a role in the development of KS, other agents must be involved in the etiology of KS. In February 2006, a new vaccine against rotavirus, RotaTeq, was licensed by the US Food and Drug Administration, and currently the Advisory Committee on Immunization Practices recommends routine immunization of all US children against rotavirus, starting at 6 weeks of age . In JuneKS is an uncommon childhood illness that is characterized by high fever and inflammation of the blood vessels throughout the body. Approximately 4,000 children each year in the United States develop KS, about eighty percent of whom are younger than 5 years of age ,4. The cBecause the etiology of KS remains unknown, and the possibility of an association between KS and rotavirus infection has been raised , we sougHospital discharge records for the period from 2000 through 2005 for California and New York residents <5 years of age with KS or with rotavirus listed as a diagnosis were analyzed from the State Inpatient Database (SID) in collaboration with the Healthcare Cost and Utilization Project (HCUP), Agency for Healthcare Research and Quality . The stath revision, Clinical Modification (ICD-9-CM) [The International Classification of Diseases, 9CD-9-CM) . For comFrom 2000 through 2005, the annual average rates of hospitalization among children <5 years of age associated with KS and with rotavirus were 20.7 and 100.7  in California, respectively, and 23.0 and 112.1 in New York, respectively.During the study period, the rate of rotavirus-associated hospitalizations displayed a characteristic winter seasonal peak with very few cases reported during the summer months Figure . With thTo further compare the seasonal pattern of hospitalizations associated with rotavirus and with KS, we examined the composite proportion of hospitalizations by month of admission over the 6-year study period figure . RotavirIn both California and New York, during 2000-2005, rotavirus-associated hospitalization rates showed a strong winter seasonality and were nearly absent during the summer months, whereas KS-associated hospitalization rates occurred year round with smaller peaks during the winter months. Furthermore, the well recognized 1-2 month delay in winter peaks of rotavirus hospitalizations in the Northeast United States compared to winter peak the West United States  was not It should be noted that although hospital discharge data are very useful in understanding disease occurrence, limitations in this methodology exist, for instance, not all rotavirus infections result in hospitalization. In addition, studies of this nature do not allow for assessment of individual-level associations between etiologic agents. Further discussions of limitations regarding this data source for KS and rotavirus, have been described elsewhere ,4,12. FiThe occurrence of KS year-round indicates that etiologies other than rotavirus are involved in the occurrence of KS. Carefully designed prospective studies with microbiologic evaluation of clinical specimens from KS cases and an appropriate comparison group with similar age, sex, and genetic background are necessary to fully evaluate any possible causal relationship between KS and infection with rotavirus.The authors declare that they have no competing interests.AM participated in the design of the study and helped draft the manuscript. RH conceived of the study, participated in the design of the study, and participated in the statistical analysis. KY participated in the statistical analysis. CS coordinated the data collection and participated in the statistical analysis. UP conceived of the study, participated in the design of the study, and helped draft the manuscript. EB conceived of the study, participated in the design of the study, and helped draft the manuscript. All authors read and approved the final manuscript.The pre-publication history for this paper can be accessed here: