Complications of suprapubic catheter insertion are rare but can be significant. We describe an unusual complication of a delayed bowel perforation following suprapubic catheter insertion.A gentleman presented with features of peritonitis and feculent discharge along a suprapubic catheter two months after insertion of the catheter.Bowel perforation is the most feared complication of suprapubic catheter insertion especially in patients with lower abdominal scar. The risk may be reduced with the use of ultrasound scan guidance. Suprapubic catheterization is a common urological procedure. Complications of catheter insertion are uncommon but can be serious including bowel perforation or obstruction. We describe an unusual complication of delayed bowel perforation after suprapubic catheter insertion.® trocar with ultra sound guidance to measure the bladder volume which was estimated as 500 ml. He had been diagnosed with poorly differentiated carcinoma of the prostate six years earlier and had undergone radical radiotherapy, bilateral sub-capsular orchidectomy and transurethral resection of prostate gland during the interim period. He had bilateral ureteric stents inserted for obstructive uropathy six months earlier. He had a past history of abdomino-perineal resection for rectal carcinoma fourteen years earlier as a curative procedure.An 86 year old gentleman had a suprapubic catheter inserted for bladder outlet obstruction. This was done under a local anaesthetic using the standard Lawrence Add-a-cathHe returned for the first change of supra pubic catheter to the Urology suite in two months time. The catheter was changed easily by the specialist nurse and the patient was discharged home. He returned about ten hours later with features of peritonitis and feculent discharge along the supra pubic catheter. He underwent an emergency explorative laparotomy. A loop of small bowel – adhered to the scar – was placed between the anterior abdominal wall and the bladder. The supra pubic tract was seen to pass through and penetrate the loop in two places before going into the urinary bladder figure . There wPerforation of the abdominal viscera is well documented as a rare but important major complication of suprapubic cystostomy ,2. To ouOur case explains the increased risk of bowel damage during suprapubic catheterization in patients with history of previous lower abdominal surgery as the bowel frequently adheres to the scar. In one study, it was found that 59% of patients with midline laparotomy incision have anterior abdominal wall adhesions . TherefoThe author(s) declare that they have no competing interests.SA Collected the data; drafted and revised the manuscript and drew the illustration. AM helped to draft and revise the manuscript and PR was the main surgeon and helped to revise the manuscript.The pre-publication history for this paper can be accessed here:


The ultraviolet properties of textiles dyed with synthetic dyes have been widely reported in literature. However, no study has investigated the ultraviolet properties of natural fabrics dyed with natural colorants. This study reports the Ultraviolet Protection Factor (UPF) of cotton fabrics dyed with colorants of plant and insect origins.Three cotton fabrics were dyed with three natural colorants. Fabrics were characterized with respect to fabric construction, weight, thickness and thread count. Influence of fabric characteristics on Ultraviolet Protection Factor was studied. Role of colorant concentration on the ultraviolet protection factor was examined via color strength analysis.A positive correlation was observed between the weight of the fabric and their UPF values. Similarly, thicker fabrics offered more protection from ultraviolet rays. Thread count appears to negatively correlate with UPF. Dyeing with natural colorants dramatically increased the protective abilities of all three fabric constructions. Additionally, within the same fabric type UPF values increased with higher depths of shade.Dyeing cotton fabrics with natural colorants increases the ultraviolet protective abilities of the fabrics and can be considered as an effective protection against ultraviolet rays. The UPF is further enhanced with colorant of dark hues and with high concentration of the colorant in the fabric. High, short-term exposure to ultraviolet radiation (UVR) from the sun causes sunburns and long-term exposure leads to skin cancer. The National Toxicology Program, U.S. Department of Health and Human Services has classified UVR as a known human carcinogen . The AmeThe ultraviolet radiation (UVR) band consists of three regions: UV-A (320 to 400 nm), UV-B (290 to 320 nm), and UV-C (200 to 290 nm). UV-C is totally absorbed by the atmosphere and does not reach the earth. UV-A causes little visible reaction on the skin but has been shown to decrease the immunological response of skin cells [Other than drastically reducing exposure to the sun, the most frequently recommended form of UV protection is the use of sunscreens, hats, and proper selection of clothing. Unfortunately, one cannot hold up a textile material to sunlight and determine how susceptible a textile is to UV rays. Even textiles which seem to be non-light transmitting may pass significant amounts of erythema-inducing UV irradiation . TherefoTo the author's knowledge, no study has investigated the ultraviolet properties of natural fabrics dyed with natural colorants. A plethora of previous studies have concluded that good UVR protection can be provided by synthetic fibers dyed with high concentrations of synthetic dyes. However, synthetic fibers such as polyester are hydrophobic and are generally not deemed to be comfortable for wear especially in warm weather. According to a report in America's Textile Industries  natural ® UV-100 F Ultraviolet Transmission Analyzer. The effect of colorant concentration on the ultraviolet protection factor was examined via color strength analysis using a HunterLab ColorQuest XE® spectrophotometer.In this study, cotton fabrics were dyed with three natural colorants of plant and insect origin. Fabrics were characterized with respect to fabric construction, weight, thickness and thread count. Ultraviolet Protection Factor (UPF) was measured using a labsphereThree fabrics were chosen to cover the gamut of basic weave constructions. They were a bleached, mercerized plain weave cotton fabric, a bleached mercerized cotton twill and a desized and bleached cotton sateen. Fabric weight was measured according to ASTM Test Method D3776-96 . Fabric Rubia tinctorum) and indigo (Indigofera tinctoria) and the natural colorant of insect origin was cochineal (Dactylopius coccus). Since natural dyes do not have affinity for cellulosic fibers an alum mordant was used to impart affinity. Fabrics were mordanted prior to dyeing by treating with alum at boil for 45 minutes. The liquor ratio was 1:40 and alum concentration was 10% on weight of the fabric. After mordanting, fabric was squeezed thoroughly and dyed.Natural plant colorants used were madder  for the skin when it is protected. The UPF is defined as the ratio of ED to EDm and calculated as follows [Direct and diffuse UV transmittance through a fabric is the crucial factor determining the UV protection of textiles . Ultravi follows :where:λ = erythemal spectral effectivenessEλ = solar spectral irradiance in Wm-2nm-1Sλ = spectral transmittance of fabricTλ = the bandwidth in nmΔλ = the wavelength in nmin vitro using a labsphere® UV-100 F Ultraviolet Transmission Analyzer according to standard AS/NZ 4399:1996 [UPF's were measured 399:1996 . Fabrics399:1996 . In no eMeasured UPF values were also correlated to the color strength of the dyed fabrics. Color strength was evaluated using K/S values generated by a HunterLab ColorQuest XE diffuse/8° spectrophotometer. K/S is a function of color depth and is represented by the equation of Kubelka and Munk (Equation 2). Higher the value of K/S greater is the color strength ,15.where R is the reflectance of the dyed fabric; K is the sorption coefficient, and S is the scattering coefficient. The spectrophotometer was standardized for a 1 inch diameter specimen viewing aperture in reflectance – specular included mode. Illuminant D65 and CIE 10-degree observer were used. During measurements, fabric samples were held flat and securely using a spring-loaded sample clamp. Three measurements were taken on each dyed fabric with the fabric rotated between measurements.2 and a thickness of 0.069 centimeters has the highest UPF value followed by the sateen weave fabric which weighed 235 g/m2 and was 0.061 centimeters thick. The plain weave fabric with a weight of 120 g/m2 and a thickness of 0.035 centimeters offers no protection against transmittance of UV rays. The positive correlation between fabric weight and fabric thickness with UPF values can be explained with reference to porosity. Porosity is a measure of tightness of weave and is also called as Coverfactor. Cover factor is defined as the percentage area occupied by warp and filling yarns in a given fabric area [Fabric characterization parameters and UPF values prior to dyeing are listed in Table ric area ,17. The ric area . The datric area . In otheThe percent UV transmittance data in the presence and absence of colorants for the plain weave fabric is shown in Figure The K/S values of the dyed fabrics which are a measure of color depth seem to support the claim that higher color depths increases UPF values. For example, in the case of the madder dyed samples when the K/S value increased from 0.20 to 0.38 the UPF values rose from 11.1 to 16.6. However, the relationship of K/S with UPF is limited to the same fabric type and the results cannot be generalized across fabrics of different weave structures. A primary reason for this observation is the acknowledgement that UPF values are dependent on a multitude of fabric construction factors such as pores in the fabric, thickness, and weight in addition to processing parameters such as dyeing and finishing. Another probable reason is the dependence of K/S on the absorbing properties of colorants in the visible region of the spectrum and that may not influence the absorption characteristics of colorants in the UV region.The percent UV transmittance data in the presence and absence of colorants for the twill weave fabric is shown in Figure Table Fabric weight and thickness are important predictors of UPF values for undyed cotton fabrics. In general, it was found that increase in weight and thickness increased the UPF though the relationship was not linear. UPF of undyed fabrics was significantly enhanced by dyeing with natural colorants especially for fabrics such as the plain weave and the sateen weave fabrics that displayed no protective abilities in the undyed state. The degree of protection imparted after dyeing was a function of the concentration of the colorant in the fabric. Within the same fabric type, as the percentage depth of shade increased so did the UPF values. In addition, darker colors such as indigo provide better protection on account of higher UV absorption. Based on the results of this study it can be theorized that plain, twill or sateen weave cotton fabrics dyed with natural colorants can provide good protection against ultraviolet rays with the only condition being that either the color has to be a dark hue or the concentration of the colorant in the fabric has to be high.The author(s) declare that they have no competing interests.AKS conceived the study, carried out the dyeing and testing and drafted the manuscript.The pre-publication history for this paper can be accessed here:


The twelve active CA isozymes are thought to regulate a variety of cellular functions including several processes in the reproductive systems.Carbonic anhydrase (CA) classically catalyses the reversible hydration of dissolved COThe present study was designed to investigate the expression of transmembrane CAs, CA IX and XII, in the mouse uterus, ovary and placenta. The expression of CA IX and XII was examined by immunoperoxidase staining method and western blotting. CA II and XIII served as positive controls since they are known to be present in the mouse reproductive tract.The data of our study indicated that CA XII is expressed in the mouse endometrium. Only very faint signal was observed in the corpus luteum of the ovary and the placenta remained mainly negative. CA IX showed weak reaction in the endometrial epithelium, while it was completely absent in the ovary and placenta.The conservation of CA XII expression in both mouse and human endometrium suggests a role for this isozyme in reproductive physiology. CAs are produced in a variety of tissues where they participate in several important biological processes such as acid-base balance, respiration, carbon dioxide and ion transport, bone resorption, ureagenesis, gluconeogenesis, lipogenesis and body fluid generation [Carbonic anhydrases (CAs) are zinc-containing metalloenzymes that are responsible for the reversible hydration of carbon dioxide in a reaction COneration ,2. The mneration ,3,4. CA neration ,6. CA VIneration . The cluneration -11. The neration ,13.It has been previously suggested that CAs may play important roles in the uterine endometrium by maintaining the appropriate pH balance through the catalysis of the production of bicarbonate ions . Indeed,CA activity has been studied in pig, horse, cow, mink, rat and human placentas, and the results show considerable heterogeneity among different species . Previouet al [CA IX is expressed at the basolateral plasma membrane of the human, rat and mouse epithelial cells ,26. In aet al  analyzedet al [Human CA XII is expressed in several organs including colon, kidney, and pancreas -30. In tet al  further In this report we studied the expression of CA II, IX, XII and XIII in mouse female genital organs including uterus, ovary and placenta. The studies were specially focused on CA IX and XII, which have been designated as tumor-associated isozymes ,10. In aIn the present study, we used the following antibodies which have been produced and characterized earlier: rabbit anti-mouse CA II , rabbit 2 asphyxiation followed by decapitation. Uterus, ovary and placenta samples were collected from both animals. The samples were immersion-fixed overnight in Carnoy's fluid (ethanol, chloroform and acetic acid (6:3:1)). Then the specimens were treated with absolute ethanol for 30 min, with 1:1 mixture of ethanol and chloroform for 15 min, and finally with chloroform for 30 min. Paraffin embedding was performed in a vacuum oven for 2 h at +58°C. Paraffin wax was purchased from Fluka Chemie GmbH (Buchs, Schwitzerland). To collect a placenta sample, a mouse was sacrificed at 9 days of pregnancy. The ninth day was chosen since it represents the middle gestational phase. It is also the time when the most critical steps of organogenesis occur in mouse. For western blotting, uterus, kidney and colon were removed and rapidly frozen in liquid nitrogen. The tissue samples for western blot were homogenized with HEPES buffer. Total protein concentration was determined after homogenization using BCA Protein Assay Kit (Pierce, Rockford, IL) according to manufacturer's instructions. The study protocols were approved by the Animal Care Committee of Tampere University.Two adult Balb/c mice were sacrificed by CO2O2) as chromogen. The sections were washed three times for 10 min in PBS after incubation steps b and d and four times for 5 min after incubation step e. All of the incubations and washings were carried out at room temperature. The sections were finally mounted in Neo-Mount (Merck, Darmstadt, Germany). The stained sections were examined and photographed with a Zeiss Axioskop 40 microscope (Carl Zeiss, Göttingen, Germany).In the mouse tissues, the localization of CA IX and XII was examined by immunoperoxidase method. Antibodies against CA II and XIII were used as positive controls for the immunostaining. All experiments were performed in duplicate and included control staining with non-immune normal rabbit serum (NRS). NRS was obtained from a rabbit that was later immunized against mouse CA XIII. The tissue samples fixed in Carnoy's fluid and embedded in paraffin were cut at 5 μm sections and placed on microscope slides. The peroxidase-anti-peroxidase complex method included the following steps: a) pretreatment of the sections with undiluted cow colostral whey (Biotop, Oulu, Finland) for 40 min and rinsing in phosphate-buffered saline (PBS); b) incubation for 1 h with the primary antiserum (anti-mouse CA II, CA IX, CA XII or CA XIII) or NRS diluted 1:100 in PBS containing 1% bovine serum albumin (BSA) (BSA-PBS solution); c) treatment with undiluted cow colostral whey (40 min); d) incubation for 1 h with swine anti-rabbit IgG (Dakopatts, Copenhagen, Denmark) diluted 1:100 in 1% BSA-PBS; e) incubation for 30 min with peroxidase-anti-peroxidase rabbit conjugate (Dakopatts) diluted 1:500 in PBS; f) incubation for 2 min with 3,3'diaminobenzidine tetrahydrochloride (DAB) solution  and Na+/H+-exhanger isoform 1 (NHE1) [It is known that CA activity facilitates transport of CO1 (NHE1) -40. EvenPrevious studies have shown CA activity in the endometrium of several mammalian species ,42. UntiIn the present study, CA XII and II showed more intense staining in the surface endometrial epithelia than CA IX and XIII. CA XII was more intensely stained in the deeper endometrial glands, while CA II was confined to the surface epithelial cells. Interestingly, CA II showed positive immunoreaction not only in the cytoplasmic compartment but also at the plasma membrane of the cells that is quite surprising for a cytosolic enzyme. The same phenomenon is detectable in some other tissues including the human gallbladder  and gut In the future studies, it will be important to investigate whether the hormonal status regulates the expression of different CA isozymes – particularly CA XII – in the endometrium. Another interesting line of investigations would be to analyze the fertilization capacity of CA XII knockout mice as soon as they become available. One could hypothesize that endometrial CA isozymes are important factors, contributing to the appropriate bicarbonate concentration and pH balance in the cervical and endometrial mucus needed for normal fertilization process. Based on our recent studies, CA IX-deficient mice showed no apparent phenotypic changes linked to fertility . Even moThe present paper demonstrates for the first time the expression of transmembrane carbonic anhydrase isozymes IX and XII in the female murine reproductive tract. The data indicates that the endometrial epithelium is a prominent site for CA XII expression. The conservation of CA XII expression in the endometrium of different species (mouse and human) suggests a role for this isozyme in reproductive physiology.All authors participated in the design of the study. PH, JL and SP collected the tissue samples. PH, JL, ET, PK and SP drafted the manuscript. PH, JL and SP performed the western blotting. SPas, JP, AW and WSS provided the antibodies. PH, JL and SP participated in the immunohistochemical staining. All authors read and approved the final manuscript.


Macaca fascicularis), a species closely related to humans that also represents a cogent animal model for the biomedical analysis of CD38.The CD38 transmembrane glycoprotein is an ADP-ribosyl cyclase that moonlights as a receptor in cells of the immune system. Both functions are independently implicated in numerous areas related to human health. This study originated from an inherent interest in studying CD38 in the cynomolgus monkey (CD38 primers, demonstrating the striking conservation of CD38 in these primates. Transfection of the cDNA coincided with: (i) surface expression of cynomolgus macaque CD38 by immunofluorescence; (ii) detection of ~42 and 84 kDa proteins by Western blot and (iii) the appearance of ecto-enzymatic activity. Monoclonal antibodies were raised against the cynomolgus CD38 ectodomain and were either species-specific or cross-reactive with human CD38, in which case they were directed against a common disulfide-requiring conformational epitope that was mapped to the C-terminal disulfide loop.A cDNA was isolated from cynomolgus macaque peripheral blood leukocytes and is predicted to encode a type II membrane protein of 301 amino acids with 92% identity to human CD38. Both RT-PCR-mediated cDNA cloning and genomic DNA PCR surveying were possible with heterologous human This multi-faceted characterization of CD38 from cynomolgus macaque demonstrates its high genetic and biochemical similarities with human CD38 while the immunological comparison adds new insights into the dominant epitopes of the primate CD38 ectodomain. These results open new prospects for the biomedical and pharmacological investigations of this receptor-enzyme. Aplysia ADPR cyclase [2+ release [Just over a decade after being identified as a leukocyte surface antigen with receptorial activity ,2, CD38  cyclase ,6, whichAplysia cyclase [Human CD38 is a broadly expressed type II transmembrane glycoprotein of ~45 kDa in its monomeric form . This co cyclase . The CD3 cyclase ,12, and  cyclase .2+ caused by the CD38/cADPR/NAADP axis has been implicated in a variety of physiological and pathological processes including insulin secretion and diabetes [The mobilization of intracellular Cadiabetes , myometrdiabetes , airway diabetes , vasculadiabetes , osteocldiabetes , and thediabetes , renal [diabetes  and exocdiabetes  systems.diabetes -24, regudiabetes -28, inhidiabetes , to indudiabetes . In addidiabetes . CD38 isdiabetes  and therdiabetes .In this study, we describe the molecular cloning and functional expression of CD38 from the cynomolgus macaque. In addition, with a panel of newly-raised mAbs, we comparatively analyse the macaque and human CD38 ECDs and identify new structural-functional characteristics of CD38 epitopes.CD38 5' and 3' untranslated regions. The 1113 base-pair (bp) insert contained an open reading frame of 906 bp , a transmembrane region (residues 22–44), and an ECD (residues 45–301) containing the signature 12-cysteine array. Alignment of the macaque and human CD38 polypeptides showed 92% identity and 94% similarity. There is complete conservation of the IC region while there are five conservative changes in the transmembrane region where macaque CD38 has one more residue than human CD38. Macaque CD38 has four potential N-linked glycosylation sites, as in human CD38; three are co-linear. Furthermore, macaque CD38 shows conservation of the 4 acidic residues (Glu148, Asp149, Asp157, Glu228) and 2 tryptophans (Trp127 and Trp191) that play a critical role in the ADP-ribosyl cyclase/cADPR hydrolase activities of human CD38 [130 is also maintained suggesting that, like human CD38, binding of ATP to this residue may lead to inhibition of the hydrolase activity [271 which is ADP-ribosylated in human CD38, causing inactivation [Activation of human peripheral blood mononuclear cells (PBMC) with phytohemagglutinin (PHA) strongly upregulates expression of CD38 in human T lymphocytes . Therefop Figure  that wasman CD38 . Lys130 activity . Likewistivation .CD38 has been characterized as a single-copy, 8-exon gene that spans ~70 kb [EcoRI, BamHI and HindIII and probed with their homologous CD38 cDNA are similar (data not shown), indicating that the structural organization of macaque and human CD38 is highly conserved. This was further demonstrated by the finding that the same primers previously used to amplify the 8 human CD38 exons [CD38 exons from macaque genomic DNA. The putative macaque exons could be perfectly aligned with their human counterparts [Human s ~70 kb ,38 and ns ~70 kb ,40. In a38 exons  also ampterparts  in numbeCD38 initiates at multiple start sites [cis-regulatory sequences, ~500 bp upstream of the initiation codon ATG were amplified by PCR from genomic DNA of cynomolgus macaque but also from chimpanzee (Pan troglodytes), to strengthen the alignment with the human CD38 promoter sequence, and numerous conserved general and immune-related potential binding sites were found . DNAs were transfected for heterologous expression in the NIH/3T3 cell line which does not express CD38 [To analyse surface expression and function of macaque CD38, the cDNA insert was subcloned into the pcDNA3.1 expression vector and a similar construct was prepared containing the human CD38 cDNA , an NAD analog which is converted by ADP-ribosyl cyclases such as CD38 to cyclic GDP ribose (cGDPR). Unlike cADPR, cGDPR is a stable, fluorescent end-product which can be detected in cell supernatants . IncreasIn human red blood cells (RBCs), CD38 is the only source of ecto-cyclase activity  which waTo establish the approximate molecular weight of macaque CD38, SDS-PAGE and Western blot analysis were performed with lysates prepared from NIH/mac38 and NIH/hum38 cells. Blots were probed with the AT1 anti-human CD38 mAb which detected a band of ~42 kDa in both transfectants Figure .To raise mouse mAbs against macaque CD38, live NIH/mac38 cells were used for immunization. Four mAbs, KK1B5 (IgG1), KK4E5 (IgG2a), KK6A11 (IgG2a) and KK9H4 (IgG1) were selected for further analyses. All four anti-macaque CD38 mAbs reacted by IF with NIH/mac38 but were negative with the parental cell line . Instead mAbs KK4E5 and KK6A11 only recognized a band of ~84 kDa, even in reducing conditions. No bands were detected by these mAbs in parental NIH/3T3 cells.Additional Western blot analyses were carried out with the anti-macaque CD38 mAbs. MAbs KK1B5 and KK9H4 confirmed detection of a ~42 kDa band in NIH/mac38 cell lysates but also recognized a second band of ~84 kDa. Figure . The douThe observation that mAbs KK4E5 and KK6A11 recognize only cynomolgus CD38 while mAbs KK1B5 and KK9H4 also recognize human CD38 indicates that the two mAb subsets are directed towards different epitopes. To evaluate the contribution of disulfides to these epitopes, mAb reactivity was assessed after treating NIH/mac38 with dithiothreitol (DTT). Reduction had no effect on binding of the species-specific mAbs (KK4E5 and KK6A11) but significantly reduced binding of mAbs KK1B5 and KK9H4 Figure , indicatReciprocal experiments were performed to see if the correlation between cross-reactivity and epitope sensitivity to DTT was also valid for a panel of 6 well-known mAbs raised against human CD38. MAbs IB4, IB6, OKT10, SUN-4B7, AT1 and HB7 were assessed for binding to native and DTT-treated NIH/hum38, and for cross-reactivity with cynomolgus CD38. Binding of mAbs IB4, IB6 and HB7 to human CD38 was unaffected by DTT and none bound cynomolgus CD38 Figure  and 4D. A priori knowledge of the human CD38 epitope map previously established that OKT10 binding is abrogated by deletion of one or both of the C-terminal Cys residues (Cys287 and Cys296) predicted to pair in disulfide bond formation [288 and Cys297 in cynomolgus CD38 (Cys287/Cys296 in human CD38).The observation that mAbs KK1B5 and KK9H4 (raised against cynomolgus CD38) and mAbs OKT10, AT1 and SUN-4B7 (raised against human CD38) all bind native but not reduced cynomolgus and human CD38 is compatible with their binding the same epitope. ormation  and thatormation . This woΔ285, a human CD38 deletion mutant which lacks the 15 C-terminal amino acids and loses OKT10 binding [Δ285 (data not shown). These data are consistent with the presence of two close conformational epitopes in human and cynomolgus CD38: one identified by mAbs KK1B5, KK9H4 and OKT10 located in the last (6th) disulfide loop, the other by mAbs SUN-4B7 and AT1 mapping to the penultimate (5th) C-terminal disulfide loop involving Cys254-Cys275  and the rhesus monkey (Macaca mulatta) (MO, FS and SC, unpublished observations).In this study, we describe the molecular cloning and functional expression of the CD38 receptor/enzyme from the cynomolgus macaque. The cDNA described here presents the expected homology to human CD38 considering that the macaque-human lineages diverged some 25 million years ago  and theiThe conceptual translation of the cynomolgus macaque CD38 cDNA yielded a polypeptide with the characteristic type II structure, size, catalytic core residues and 12-cysteine ECD array common to CD38 orthologs. With respect to human CD38, cynomolgus CD38 has an extra residue in the transmembrane domain but no difference was found in the IC tail, where human CD38 is reported to interact with the SH2 domain of Lck  in lipidCross-reactivity of anti-human CD38 mAbs was exploited in the initial part of the protein analyses although these give discrepant results in binding to leukocytes from other primates. For example, OKT10, but not Leu17 or T16, binds bone marrow from rhesus macaques  whereas N-linked glycosylation motifs, suggesting it is probably glycosylated, like human CD38 [The apparent molecular weight of macaque and human CD38 were indistinguishable by SDS-PAGE. The macaque polypeptide weighs 34.4 kDa and has four man CD38 , to giveman CD38 ,51, non-man CD38  and tranman CD38 . Therefoman CD38 , or a trThe availability of the macaque CD38 amino acid sequence and its alignment with the human homolog gave a new dimension to the analysis of mAb cross-reactivity and ultimately led to a better understanding of CD38 epitopes. Macaque and human CD38 are strikingly conserved yet only two of the four anti-macaque CD38 mAbs cross-reacted, as did only three of the six anti-human CD38 mAbs in reciprocal experiments. The key finding was that a mAb's capacity to cross-react always correlated with the sensitivity of its target to reduction which, added to the prior knowledge of the human CD38 epitope map, crystal structure and active site , allowede.g., detection of membrane CD38 in apoptotic cells might be positive according to type I mAbs and negative by type II . Secondly, simultaneous use of the two types of mAbs can provide information about CD38 expression (type I) while the type II mAb can give indications on its conformation. Thirdly, it is possible that potent Ca2+-mobilizing agonistic mAbs are more likely to be type I mAbs since this subgroup includes IB4, which is the only anti-human CD38 mAb to mobilize Ca2+, and NIM-R5, a rat anti-murine CD38 mAb [2+ and whose binding to murine CD38 transfectants was not affected by DTT . Finally, autoantibodies to CD38 have been detected in diabetes and thyroiditis and it would be interesting to identify the epitopic culprits.The classification of anti-primate CD38 mAbs as species-specific/DTT-resistant (type I) or cross-reactive/DTT-sensitive (type II) and directed against a conformational epitope may be of practical importance. Firstly, our results demonstrate the necessity for careful antibody selection when performing biological assays involving detection of CD38 expression in circumstances of cell membrane redox perturbation, CD38 mAb  which alMacaca fascicularis have been elucidated and new insights obtained about the epitopic structure of the CD38 ECD. We hope that, by providing the reagents for analysis of CD38 in the cynomolgus macaque, this study may expedite our understanding of the role of CD38 in human disease.Some of the essential biological features of CD38 in PBMC were isolated by Ficoll-Paque PLUS (Amersham Biosciences) centrifugation from whole blood obtained from a female cynomolgus macaque housed according to state regulations at the RBM . Cells were cultured in medium with 5 μg/ml PHA (Sigma-Aldrich) for 72 h, harvested and resuspended in TRIzol RNA extraction solution (Invitrogen). RT-PCR was carried out using the Titan One Tube RT-PCR system (Roche Diagnostics) with 150 ng total RNA and the gene-specific primers derived from the human CD38 sequence (GenBank accession no. D84278): forward 5'-AGT TTC AGA ACC CAG CCA-3' (corresponding to nt -84 to -66 upstream of the ATG initiation codon); reverse 5'-ATT GAC CTT ATT GTG GAG G-3' (corresponding to nt 102–121 downstream of the TGA stop codon). After RT for 50°C for 30 min, the cDNA was amplified by two rounds of PCR: 5 min at 94°C, followed by 30 (first round) or 35 (second round) cycles at 94°C for 30 s, 54°C for 30 s and 2 min at 72°C. The product was gel purified and cloned into the pGEM-T Easy vector (Promega). The inserts of three recombinant clones were analyzed by automated sequencing (Applied Biosystems).CD38 exons [CD38 promoter of cynomolgus macaque was amplified  with a forward primer from the human CD38 promoter (5'-GAA GAG GCA AGA AAA GCC-3') and reverse primer chosen from macaque CD38 exon 1 (5'-AACTCG CAG TTG GCC ATA-3'). The chimpanzee CD38 promoter was amplified with the human CD38 sequences. The 5' end of cynomolgus CD38 intron 1 was amplified  with forward primer 5'-CCG TCC TGG CAC GAT GCG TCA AG-3' from macaque exon 1, and reverse primer 5'-ACA CCC TCC TCC CCT ACC ACA GG-3' taken from human CD38 intron 1. Amplicons were gel purified and analysed by automated sequencing. Alignments were performed with CLUSTALW (ExPASy, Swiss Institute of Bioinformatics).Cynomolgus macaque (15 samples) and chimpanzee (5 samples) genomic DNAs were obtained as described . The eig38 exons  while thΔ285 plasmid [Δ285 transfectant was kindly provided by Dr. Umberto Dianzani  [2 goat anti-mouse Ig-FITC was used as secondary antibody (Jackson ImmunoResearch Laboratories).NIH/3T3 murine fibroblast and COS-7 monkey kidney cell lines were from the ATCC. Production of SL-691 and SL-999 cynomolgus B lymphoblastoid cell lines was previously described . The mut plasmid  was kind, Italy) . Cells wcDNAs were cloned into pcDNA3.1/V5-His-TOPO expression vector (Invitrogen). Stable transfected cell lines were produced in NIH/3T3 by electroporation (250 V, 960 μF at 20°C), selected for 3–4 weeks in G418 after which isolated clones were picked and transferred to 96-well plates.5 cells/ml in PBS. For RBCs, 420 μl packed volume were brought to 1 ml by addition of NGT buffer (0.15 M NaCl, 5 mM glucose, 10 mM Tris. Cl, pH 7.4). To 1 ml cell suspensions 10 μl 10 mM NGD (Sigma) in 20 mM Tris, pH 7.4 or 10 μl buffer (control) were added. After 30 min at 37°C, supernatants were collected after brief centrifugation. Supernatants were analysed by fluorescence spectrometer set at excitation wavelength 300 nm and emission wavelength 410 nm. Cell lines were measured in triplicate in three independent experiments; RBCs from two different animals were tested once; RBCs from humans were tested in duplicate in two independent experiments.Ecto-GDPR cyclase activity of intact cells was determined as previously described ,30,61. BCells were lysed in NP-40 buffer (150 mM NaCl, 1.0% NP-40, 50 mM Tris pH 8.0) containing protease inhibitors. Samples (20 μg protein/lane) were analysed by 8 or 10% SDS-PAGE and transferred to PVDF membranes (Bio-Rad Laboratories, Hercules, CA). Membranes were blocked in 5% milk/TBST, incubated for 2–3 h at RT with mAb supernatant with 1% milk, and incubated with horseradish peroxidase-labeled anti-mouse IgG (PerkinElmer) followed by ECL visualization.5 live NIH/mac38 cells in PBS. Eight days later, mice received an i.p. boost of NIH/mac38 cells, and mouse sera tested 8 days later. The spleen of one mouse that responded to immunization was selected for fusion to P3.X63.Ag8.653 murine myeloma cell line. Hybridoma supernatants were screened for reactivity with the immunizing cells and lack of reactivity with the parental cell line. Positive hybridomas were cloned by limiting dilution. MAbs were used in the form of supernatants containing NaN3.BALB/c mice (Charles River Laboratories) were anaesthetized with Avertin i.p. injection and immunized by intrasplenic injection with 300 μl containing 5 × 105 cells/sample were incubated with 100 μl mAb supernatant/NaN3 for 1 h at 4°C, and 30 min at 4°C with FITC-labeled secondary Ab. Background fluorescence was established with control IgG antibody. Cells were analysed immediately either by fluorescence microscope, FACSCalibur flow cytometer (10,000 events acquired) with CellQuest software (Becton Dickinson) or Olympus FV3000 confocal microscope with Nomarski optics for differential interference contrast (DIC) and FluoView 300 software.Surface expression of cynomolgus and human CD38 was determined by IF. Briefly, 2 × 106 cells/ml. One hundred μl 100 mM DTT stock was added per ml cell suspension, kept for 45 min in a 37°C incubator and washed twice with PBS/BSA/NaN3 before analysis.Cells were detached with 1 mM EDTA/PBS, washed and resuspended in complete medium at a concentration of 2 × 10EF carried out cellular, biochemical and enzymatic analyses, devised the comparative epitope mapping and wrote the manuscript; MO cloned the cDNA and participated in the genomic DNA analyses; PV carried out mAb production and confocal microscopy, and participated in cellular and biochemical assays; EO carried out FACS analyses; SC participated in the genomic DNA analyses and contributed primate DNA samples; FS designed and participated in the cDNA and genomic DNA analyses; FT carried out expression analyses in macaques; FM designed and supervised mAb generation, and edited the manuscript. All authors read and approved the final manuscript.


It's a basic biological principle that living things share certain fundamental traits. That's why understanding the mechanisms of cell division in single-celled yeast, say, can offer insight into cell division in humans. Now Joseph Heitman and colleagues report that the evolutionary events that spawned sex chromosomes in yeast resemble those that shaped sex chromosomes in animals.Cryptococcus, Heitman and colleagues found that this fungal sex-determining region arose via a series of discrete events that echo those that shaped mammalian sex chromosomes.Strictly speaking, yeast—the common name for single-celled fungi—don't have sex chromosomes; they have sex-determining regions within chromosomes, called mating type, or MAT, loci. In a comparative genomic analysis of the MAT locus in three species of the human pathogenic fungus a, for example, and mating type alpha. Still, yeast manage a measure of complexity and considerable elegance in the systems they deploy to sexually reproduce.A primary benefit of sexual reproduction is the genetic diversity gained from reshuffling genetic material during meiosis, which creates gametes. Yeast sex, such as it is, accomplishes the same thing. Of course, sexual identity for a fungus does not take the form of sperm or egg but of mating type a or alpha mating status are alleles (variants) of a single MAT locus. Cells with the MATa allele are mating type a, while cells with the MATalpha allele exhibit mating type alpha. A cell can switch its mating type when genetic exchange, or recombination, between two mating loci occurs.In ascomycetes, like baker's yeast, the MAT locus is small and includes just a few genes. The genes that determine a cell's Ustilago maydis—a maize pathogen that some consider a culinary delicacy—mating is more complex, and sexual identity is determined by two unlinked genomic regions with distinct classes of genes. Cells must be of different mating types at both loci to allow sexual reproduction. To their surprise, Heitman and colleagues discovered that the mating locus of Cryptococcus neoformans—a basidiomycete fungus that infects humans and is associated with transplant recipients, patients with AIDS, and other immune-compromised patients—exhibits several unique features, common to neither ascomycetes or their basidiomycete relatives.In basidiomycetes, like the corn smut C. neoformans locus occupies a single region and is unusually large, spanning more than 100 kilobases and containing over 20 genes, including those typically segregated in separate locations in other basidiomycetes. Like on the human Y chromosome, the sex-determining genes of C. neoformans are interspersed with non-sex-related genes. And unlike ascomycetes, which also have a single active MAT locus and two mating types, no mating type switching occurs as there are no silent mating type cassettes in the genome.Unlike most basidiomycetes, the a and alpha alleles of C. neoformans' closest relative, C. gattii, and compared these variants to four already characterized variants derived from two C. neoformans subspecies. All six MAT alleles share characteristic features, including a fairly large size, a common gene set, and dramatic genomic migration during evolution . Each MAT allele has genes with different evolutionary histories, ranging from ancient to recent, that fall into distinct patterns based on shared nucleotide composition and mating type. The patterns correlate with how long the genes have occupied the MAT locus, suggesting how it evolved.Heitman and colleagues sequenced the Cryptococcus MAT locus resembles the evolution and structure proposed for the ancient Y chromosome, the authors argue that Cryptococcus can serve as a valuable model to study the molecular dynamics of sex chromosomes.The authors hypothesize that this novel structure was formed by chromosomal rearrangements that linked two unrelated genomic regions into a single region. Recombination between these sex-determining regions was suppressed after other events blurred their boundaries. Specific genes in the once separated loci then attracted mobile elements in the genome to their sites, thus precipitating expansion of the locus. Because the


To examine the enabling and constraining factors that influence African social scientists involvement in malaria control.Convenience and snowball sampling was used to identify participants. Data collection was conducted in two phases: a mailed survey was followed by in-depth phone interviews with selected individuals chosen from the survey.Most participants did not necessarily seek malaria as a career path. Having a mentor who provided research and training opportunities, and developing strong technical skills in malaria control and grant or proposal writing facilitated career opportunities in malaria. A paucity of jobs and funding and inadequate technical skills in malaria limited the type and number of opportunities available to social scientists in malaria control.Understanding the factors that influence job satisfaction, recruitment and retention in malaria control is necessary for better integration of social scientists into malaria control. However, given the wide array of skills that social scientists have and the variety of deadly diseases competing for attention in Sub Saharan Africa, it might be more cost effective to employ social scientists to work broadly on issues common to communicable diseases in general rather than solely on malaria. Malaria control remains ineffective in many endemic areas in spite of efficacious interventions, such as combined antimalarial therapies and insecticide-treated materials. Biological, environmental, political, socio-cultural, economic and behavioural factors influence the transmission of malaria, thus requiring a multidisciplinary and integrated approach to effectively control the spread of malaria ,2.In recent years international initiatives, such as Roll Back Malaria, have highlighted the contributions that social scientists bring to a multidisciplinary approach for malaria control. Various studies in Africa have illustrated the benefits of collaborative research between social and biomedical scientists -17. HoweSeveral factors continue to limit the application of social science research in tropical public health in general, and malaria control in particular. These include: confusion among many non-social scientists and social scientists alike about the variations in social science theory and methods and the different types of contributions that each can make to public health and malaria control activities, a perception among many public health and malaria control professionals that the problems of communicable diseases can be solved with technical and clinical interventions, ignoring the social nature of illness, limited communication among social scientists working in applied health and malaria control, insufficient access to current social science and malaria literature, and the lack of trained social scientists with applied experience in the endemic countries ,29.Given the poor academic and professional structures found in many African countries, an increasing number of highly skilled citizens emigrate abroad for further studies but fail to return to their respective country of origin, a phenomenon commonly referred to as 'brain drain' -36. As aA convenience sample was derived from existing social science networks such as, the Partnership for Social Science and Malaria Control (PSSMC) Network, the Social Science and Medicine Africa Network (SOMA-Net), Pan African Anthropological Association (PAAA), as well as the CHANGE Project database of African social scientists, and the United Nations Development Program, World Bank and World Health Organization's Special Programme for Research and Training (WHO/TDR) database of social scientists trained in the last decade (1992–2000). Snowball sampling was used to elicit names of additional social scientists known personally to the participants and investigators.Social science was broadly defined to include disciplines such as anthropology, sociology, health economics, demography and population studies, development studies and public health. Enrolment criteria included specific social science training within these disciplines, and participants had to be of African origin. Enabling factors were defined as those factors that made it easy to work in malaria control, including factors that first attracted social scientists and those that facilitated entrance into the field. Constraining factors were defined as those factors that impeded work in malaria control and factors that made malaria control unattractive to social scientists.Data collection was organized in two phases. In Phase I, consent forms and questionnaires were sent by email and/or fax to potential participants. Participants could respond directly and/or refer other eligible colleagues. Those who were ineligible or uninterested could decline. For those emails that could not be delivered, three attempts were made to resend the survey and consent form. The questionnaire focused on demographic characteristics, work experiences, factors influencing career choices and sources of career development information. The survey instrument was piloted among five participants from different countries. Based on the feedback from the pilot survey, minor adjustments were made to improve use via email. Email communication was preferred but, in the absence of email, other methods of communication were used. If the survey was not returned within three weeks, attempts were made to follow-up.The survey responses in Phase I provided an overview of the involvement of the participants in malaria related activities but gave only a limited understanding of the specific factors affecting career trajectories. In Phase II, we conducted in-depth phone interviews using a purposive sample derived from Phase I that was selected according to country of origin, sex, academic field  and interest in malaria and applied or operational work experience . A deliberate effort was made to ensure that the sample represented those with or without an interest in malaria, those who actively sought employment in malaria and those who did not.The more descriptive data from Phase II clarified Phase I data. For example, with regards to participants' interest in malaria, some participants in Phase I indicated an interest in malaria while in school, but did not include malaria in their thesis research and did not pursue employment in malaria. The in-depth, open-ended phone interviews in Phase II provided greater detail as to other factors that might have influenced participants' career decisions in malaria control. Open-ended questions focused on key events and times in the career trajectory during which critical decisions were made, current and past work experiences, reasons for selecting specific jobs, technical experiences with proposal writing, challenges and rewards of working in malaria, and suggestions for improving the integration of social science in malaria control.Descriptive statistics were generated using EPI-Info. Qualitative data were transcribed and content analysis was used to develop contextual themes. The study was reviewed and approved by the Institutional Review Board of the Centers for Disease Control and Prevention. Verbatim quotes from the participants are used throughout the text to illustrate contextual points of discussion.This study was conducted between May 2002–May 2003. Of the 136 surveys sent out, 40 completed surveys were found to fit our enrolment criteria  were male between the ages of 30–39 years old Table . Fifty pOf the 18 participants selected for a phone interview, 55% (n = 10) were between 30–39 years and 22% (n = 4) were female. Of those with postgraduate degrees, 50% (n = 9) had a masters degree, 44% (n = 8) either had a doctoral degree or where doctoral degree students while 5% (n = 1) had a bachelor's degree. When difficulties arose in ascertaining whether the professional discipline linked to the social sciences, participants were contacted for clarification.Eighty-five percent of the participants (34/40) received funding for postgraduate training from their national government, international organizations (such as the WHO or DFID), or other foreign governments. Sources of career development information included the Internet, friends and colleagues, local universities, conferences, journals, professional networks and local newspapers.When asked if participants were interested in malaria during their academic training, 82% (33/40) of the participants said yes. Only 36% (12/33) of those interested included malaria as a research topic in their thesis or dissertation. Three participants (7.5%) who were not interested in malaria included malaria in their thesis. Post graduation, 35% (14/40) of the participants actively sought employment in malaria control. Three of the 14 (21%) who sought employment in malaria were turned down for jobs due to insufficient technical knowledge of malaria control, while six participants (15%) returned to previous health care positions, some of which were positions in malaria control. Although some participants were not currently working on malaria-related projects, nearly all participants, at some point in their career, had worked as a consultant in malaria control in addition to their full time positions. Participants who specialized in malaria for postgraduate degrees were often funded by a national or international organization, on condition that they would return to their jobs in malaria following the completion of studies."Malaria is an endemic disease in my community, so I can't say I was never interested. I wanted to do a thesis on malaria, but I realized that social scientists could not get jobs in malaria so I had to look elsewhere." When followed up during the phone interviews in Phase II, it was clear that some participants turned down jobs in malaria due to competing employment in areas such as HIV/AIDS, while others declined malaria control contracts due to the competing demands of doctoral-level training.Eleven of the 40 participants said that the primary factor that prevented those with an interest in malaria from seeking further research and employment opportunities in malaria control was the perception that social scientists could not be employed in malaria control. As one participant remarked "For years I thought the field of malaria was boring and I was not interested in it. I only became interested in malaria when I realized that it was a complex disease with a major public health impact and it appeared that funding for research might be available."Another issue that was revealed in Phase I was the influence of the epidemiology of malaria on participants' interest in malaria. Eleven of the 18 participants during the phone interview explained that they developed an interest in malaria after learning about the complex nature of the disease, its high prevalence and the possibility of access to research funds. All 40 participants were asked to identify factors that attracted social scientists into general employment opportunities, as well as malaria-specific opportunities Table . ResponsOpportunistic events also shaped career development. For example, the successful completion of a project, fellowship or consultancy often produced a cascade effect by opening up other opportunities in malaria control, resulting in individual professional recognition by local and international collaborators as an expert in the field. Attending workshops sponsored by international organizations, such as the World Bank and WHO, enabled participants to get their name in the organizations' database for possible consultancies."Most of the rejected proposals do not give reasons for rejection, which is unfortunate. I have been lucky to have one rejected proposal that was returned with explanations as to why it was rejected. I was told the issue we raised was very broad and vague and it made me realize that I had to be very clear in my framework. After that rejection, I was able to develop projects that had a clear framework and include a multidisciplinary team to strengthen other key areas and the next proposal we developed was accepted."Strong writing skills were identified as essential for job and training opportunities by most of the 18 participants in Phase II. Ten of the 18 participants (55%) in the phone interview noted that while it was initially difficult to identify and develop fundable proposals, receiving constructive feedback from rejected proposals helped improve their writing skills and led to the development of more successful proposals. "In Africa we take what we get. One rarely ends up in a profession of their choice. People take the job that is at hand and learn to deliver as best as they can." This was true for both those interested in malaria originally and those not interested. During the in-depth interviews, we also asked participants who did not initially express an interest in malaria but sought and gained malaria related employment, why they choose to stay in malaria? Of the 18 participants eight agreed with the following view: "I would probably stay in malaria because it is a major problem in my country, and I feel I have gained significant experience in malaria over the years. This has strengthened my confidence and ability to handle current and hopefully future opportunities, and maybe I am too old right now to start learning something new."When asked why they chose employment in malaria, one of the participants echoed many others by saying, "specifically focusing on malaria as the only area for job opportunities narrows ones fields for job opportunities. Although malaria is an endemic disease in my country, it is largely dominated by physicians and there are not sufficient job opportunities for social scientists in my country so it is impossible to focus only on malaria. You need something to fall back on besides malaria." Participants also noted that a paucity of research centres and limited funding for social science research also contributed to the lack of job opportunities. In situations where job opportunities existed for social scientists, they were generally limited to short-term rather than long-term career paths.Participants were also asked to identify factors that constrained employment in malaria. All 40 participants were more likely to refer to employment challenges specific to malaria research, rather than malaria control, as national malaria control programmes employ very few social scientists. Ten of the 18 participants in Phase II noted that social science involvement in malaria control was a recent development in their countries, as biomedical scientists had previously dominated malaria control. Another phone interview participant remarked that "At first, I did not realize the impact of malaria in my community. Everybody was talking about HIV/AIDS. I think it is because people have lived with malaria for so long that they treat it at home. It was only after doing some training at the hospital , that I realized how many people, especially children and mothers, die of malaria and I got interested in working in malaria control."Individual perceptions of malaria as a 'normal' everyday event limited participants' understanding of the impact of malaria and, thus, diminished their interest in malaria control. As one of the phone interviewed participants remarked, "I prefer working on maternal and child health issues, as well as HIV/AIDS, because personnel in these areas are able to recognize the role of social science and request social science input.  This makes me confident that my discipline is important and I can make a contribution." Of the 40 participants in Phase I, 36 identified the lack of professional development opportunities as a constraining factor to employment in malaria.  The lack of a supportive work environment with good communication and mutual respect was mentioned by those working primarily with biomedical scientists.  "Research is largely uncoordinated.  We have no knowledge of who is doing what, there is no database of social scientists and the projects that they're working on from which we can develop collaborative projects.  We neither have the infrastructure nor the access to current literature, which makes it difficult to develop manuscripts for publication."  Insufficient technical skills in proposal writing and professional isolation from other social scientists hampered the development of collaborative relationships, which made it difficult to develop competitive proposals for funding and publication.  As a participant observed, "try to learn not only about social science in malaria control, but all other aspects of malaria control as well, such as entomology, vector control and treatment. Having only a social science background without any understanding of the other areas of malaria control might make it difficult to effectively apply the social science aspect within malaria control. Besides you will always be working as part of a multidisciplinary team, therefore, you will need to understand their language in order to be able to effectively communicate the social science dimension to them."In order to meet the challenges of employment, eight of the 18 participants in Phase II stressed the importance of understanding all technical aspects of malaria control. One participant echoed the view of several others in their advice to junior social scientists, "Encourage donors to develop grants for student researchers working with senior scientists, and identify conferences where students could present research findings."Participants also emphasized the importance of linking students to practical training opportunities in endemic areas for fieldwork experience and possible long-term opportunities. As one participant noted, Participants in Phase II were asked to describe strategies for improving the integration of social science in malaria control. Of the 18 participants interviewed, 13 (72%) stressed the need for social science advocacy as an important step in malaria research and control. Advocacy efforts were recommended for social scientists and non-social scientists alike, as well as policy makers in health and education and those who fund research. Five participants said their involvement in malaria projects was as a result of specific requests for social scientists in the call for proposals.All 18 participants emphasized the need to develop outreach efforts for junior social scientists in order to enhance their understanding of the potential contributions that social scientists could offer, the type of training needed, and the types of jobs they could perform in malaria control. Three of the 18 participants also suggested the revision of graduate and medical school curricula to include malaria and social science courses, but noted that support had to come from the national level for curricula change to take place.The establishment of a sustainable network of African social scientists working in malaria control was identified as a useful tool for integrating social sciences in malaria control by six of the 18 participants. Possible functions of the network were described as developing a mentoring programme for junior social scientists, a forum to disseminate current local social science research, information relating to employment and training and funding opportunities."Make the research process more participatory. Involve policy makers and consult with them so that eventually they are the ones who demand the research and in this way, social science research can influence policy. We developed working papers with key findings for distribution to various stakeholders. We published our findings on the Internet and conducted department seminars or short courses during which we discussed our research and presented the information in a very manageable manner. At least, in this way, we were able to get our findings into the hands of policy makers and hopefully begin a dialogue with them."To facilitate the application of social science research findings to national malaria control policies, one of the phone-interviewed participants explained how to include policy makers in the process of social science research. Results of this study indicate that factors such as having an interest in malaria, a mentor during one's academic and professional career, strong writing skills, and technical experience in malaria made it possible for participants to take advantage of opportunistic events and job opportunities that led to employment in malaria control activities. As well, limited job opportunities, the lack of career development opportunities, and the lack of understanding about social scientists' role in malaria limited the involvement of social scientists in malaria control. This study revealed that many of the enabling and constraining factors for social scientists' involvement in malaria control are not unique to malaria control but are common to African researchers in general. An examination of the intrinsic and extrinsic factors regarding career trajectories of social scientists in this study revealed that the majority of participants did not specifically seek malaria as a career path but, rather, were looking for employment opportunities in general, some of which happened to be in malaria. It is unclear to what extent the career development process in many African countries contributes to the way employment is sought or if this is due to the relatively new role that social science plays in malaria control, which was noted by numerous participants. Career development is a process that ideally begins during one's formal academic training and continues throughout one's professional life. However, if career development is concentrated more towards academic training and less on postgraduate employment opportunities, it is no wonder that post graduation, students are forced to 'take whatever they can get,' as opposed to seeking opportunities that complement their interest and training.The lack of proper career counselling could also limit a student's understanding of what courses and practical experiences are most beneficial for social science and health careers, without which social scientists cannot be hired as part of a health intervention team ,41. MoreIn order to effectively design and implement malaria control interventions, it is necessary to have social scientists that are knowledgeable about the technical issues that underlie malaria control. This knowledge is acquired over time and needs to be supplemented with direct field experiences. However, given the economic constraints facing national malaria control programmes and Ministries of Health, this study shows that it might not be reasonable to expect that social scientists will be assigned solely to malaria control. Given the limited number of trained social scientists with practical experience and the variety of deadly diseases competing for attention in sub-Saharan Africa, there are few social scientists that are both willing and able to dedicate their expertise only to malaria control.Ministries of Health and Education should work with social scientists to establish employment opportunities in communicable diseases. Given the wide array of skills that social scientists have, it would be cost effective to employ social scientists to work broadly on issues common to communicable diseases, rather than solely on malaria. This "generic" approach would also support the programmatic shift from malaria as a vertical programme to its being integrated within primary health services or the wider arena of communicable diseases. Although this may not be ideal for building strong social science capacity specifically for malaria, this offers a pragmatic approach to utilizing scarce resources. Lessons learned from behavioural interventions in other diseases can also be applied to malaria control, thus strengthening social science contributions to malaria control.The acknowledgement by many participants of their need for a better understanding of both biomedical and technical aspects of malaria control is reinforced by data from other successful projects that are aimed at establishing collaborative multidisciplinary research teams. For example, the International Clinical Epidemiology Network (INCLEN) social science programme seeks to enhance clinicians' understanding of social science by exposure to relevant issues in social science design and measurement and evaluation issues, while teaching principles of clinical epidemiology and biostatistics to established social scientists. As a result, strong collaborative partnerships among individual clinical epidemiologists, social scientists, and biostatisticians have been developed to produce the interdisciplinary solutions required for priority health problems in society .A key factor identified as important for employment in malaria was the ability to contribute social science solutions to malaria control and the integration of research findings into malaria control programmes and policies. Social scientists are more likely to work directly with members of the community therefore, it is important that strong relationships are developed with local communities and their leaders, as a trust building mechanism and a matter of social prestige for the researcher. This link to the community facilitates the application of research findings on a local level ,42.While findings from this study give us greater insight into factors that affect career trajectories of African social scientists, caution must be used when interpreting the results. The use of convenience sampling and snowball sampling techniques to identify eligible participants appeared to have excluded the younger more recently qualified social scientists who might not have been members of the various networks we used to identify eligible participants for this study. It is possible that some of the junior social scientists do not have much work experience in malaria and are not yet known by the more experienced social scientists that helped with referring other participants for the study. In addition, although telephone interviewing enabled us to reach social scientists from several countries, we were not able to reach those social scientists with no access to the Internet and telephone who might have had other unique challenges not captured in this study. Further research should examine younger or more recent social science graduates as well as those with no access to the Internet.Malaria control is at a point where there is good evidence-based data to demonstrate that various approaches can make a difference in malaria-related morbidity and mortality. Prompt and timely treatments with an effective drug, intermittent preventive treatment for malaria in pregnancy, and use of insecticide-treated materials are examples of interventions that have been shown to be effective. However, the success of these approaches depends, to a great extent, on human behaviour. Epidemiologists and clinicians, who work in malaria control, traditionally, do not have the necessary training to incorporate behavioural and social science knowledge into the development, implementation, and evaluation of control interventions. It is time to broaden the perspective of malaria control, which can be best achieved through employing a multidisciplinary team that includes social scientists. To support such collaboration, attention needs to be given to understanding the factors that influence job satisfaction, recruitment and retention of the social scientists.At this point in time, social science capacity is severely limited in most African malarious areas due to insufficient funding, lack of career structure, loss of trained individuals to international postings, and a limited understanding of how social scientists can contribute to malaria control. In order to improve this situation, capacity development is needed as a first step. Joint collaborative efforts should be made to offer technical malaria knowledge to social scientists and social science methodology to epidemiologists and control personnel. Social scientists need forums and networks to exchange information, learn about existing research and educational opportunities and promote collaborative partnerships with national malaria control programmes. However, these mechanisms cannot be realized without adequate levels of funding.In order to attract skilled social scientists to Ministries of Health, a new line of thinking needs to emerge that situates social scientists more broadly within the ministry. Rather than focusing on how to integrate social scientists solely within malaria, the emphasis should shift to thinking creatively about how to best utilize social science expertise across an array of specific programmatic areas. Novel approaches within malaria control interventions are starting to emerge, such as integrating the distribution of insecticide-treated nets with measles immunizations. This same type of non-traditional thinking should be applied when developing career tracks for social scientists within Ministries of Health.However, incentives for employment must rival or exceed those offered in other areas. Pay scales must be equitable and the value of multidisciplinary perspectives must be appreciated and desired by national malaria control programmes and Ministries of Health in order for career structures for long-term employment of social scientists to be established. For contractual employment, the same considerations should be shown. Short-term salaries for malaria-related projects should be competitive with salaries offered for similar work being conducted with other diseases, such as HIV/AIDS.In the last five years, there has been a noticeable increase in the collaboration among social scientists and others engaged in malaria control. The interest in malaria by social scientists is clearly present. What are needed now are structures for training and employment that offer a professional career path over time.HAW and CJ obtained funding for the project. All authors contributed to the study conception and design. PN coordinated fieldwork, data collection and analysis with ongoing supervision from HAW and CJ. PN wrote the first draft of the article with critical revisions from HAW and CJ. PN, HAW, CJ, IN, SD and FG all read and approved the final manuscript.


The purpose of this study was to analyze pretreatment predictors of short-term weight loss in Portuguese overweight and obese women involved in a weight management program. Behavioral and psychosocial predictors were selected a priori from previous results reported in American women who participated in a similar program.2) who participated in a 4-month lifestyle weight loss program consisting of group-based behavior therapy to improve diet and increase physical activity. At baseline, all women completed a comprehensive behavioral and psychosocial battery, in standardized conditions.Subjects were 140 healthy overweight/obese women  did not finish the program. By treatment's end, more than half of all women had met the recomended weight loss goals, despite a large variability in individual results (range for weight loss = 19 kg). In bivariate and multivariate correlation/regression analysis fewer previous diets and weight outcome evaluations, and to a lesser extent self-motivation and body image were significant and independent predictors of weight reduction, before and after adjustment for baseline weight. A negative and slightly curvilinear relationship best described the association between outcome evaluations and weight change, revealing that persons with very accepting evaluations  lost the least amount of weight while positive but moderate evaluations of outcomes (i.e., neither low nor extremely demanding) were more predictive of success. Among those subjects who reported having initiated more than 3–4 diets in the year before the study, very few were found to be in the most successful group after treatment. Quality of life, self-esteem, and exercise variables did not predict outcomes.Several variables were confirmed as predictors of success in short-term weight loss and can be used in future hypothesis-testing studies and as a part of more evolved prediction models. Previous dieting, and pretreatment self-motivation and body image are associated with subsequent weight loss, in agreement with earlier findings in previous samples. Weight outcome evaluations appear to display a more complex relationship with treatment results and culture-specific factors may be useful in explaining this pattern of association. Predicting weight loss outcomes from information collected from subjects before they start weight management programs is a long-standing goal . In effeAmong the most valuable applications of valid weight loss prediction models is the early identification of individuals with the least estimated probability of success in a given treatment, who could (and perhaps should) be directed to alternative therapies. Research specifically aimed at studying these overweight/obese persons, who are more resistant to current forms of treatment, would be particularly relevant. Equally important are improvements in the matching between treatments and participants, which are dependent on the measurement of relevant pretreatment variables (i.e., that are found to predict success). More individualized programs have the potential for higher cost-effectiveness and improved overall success rates, by targeting specific areas of concern in selected participants or homogeneous groups . FinallyWe have previously tested a large number of psychosocial and behavioral variables as predictors of short-term weight outcomes . A numbe2, and be free from major disease to be eligible for the study. After several orientation sessions, 152 women signed up for the program. During the run-in phase, four women decided not to participate (reporting new time and scheduling conflicts), four did not comply with testing requirements and were excluded, three women found out they were pregnant or decided to attempt pregnancy and were also excluded, and one subject was found ineligible due to medical reasons (untreated hyperthyroidism), leaving a total of 140 women who started the intervention. An initial visit with the study physician ensured that subjects met all medical inclusion criteria. All participants agreed to refrain from participating in any other weight loss program and gave written informed consent prior to participation in the study. The Faculty of Human Movement's Human Subjects Institutional Review Board approved the study.Subjects were recruited from the community for a 2-year weight management program through newspaper ads, a website, email messages on listservs, and announcement flyers. Subjects were required to be older than 24 years, be premenopausal and not currently pregnant, have a BMI higher than 24.9 kg/mWeight was measured twice, to the nearest 0.1 kg (average was used) using an electronic scale (SECA model 770, Hamburg, Germany) and height was also measured twice, to the nearest 0.1 cm (average was used). Body mass index (BMI) in kilograms per squared meter was calculated from weight (kg) and height (m). In addition to weight and other morphological and physiological variables assessed, subjects filled out a large psychosocial questionnaire battery prior to the first weekly treatment session. This was conducted in standardized conditions of comfort and silence, with a study technician attending every assessment period. To ensure optimal levels of concentration and avoid overburden caused by long periods of psychometric testing, subjects were required to attend three sessions, each lasting approximately 45 minutes.Portuguese versions of the Impact of Weight on Quality of Life – Lite (IWQOL-Lite, ), Self-MNumber of previous diets and weight history variables were taken from a diet/weight history questionnaire developed specifically for this study. Weight outcome evaluations were assessed by 4 questions derived from the Goals and Relative Weights Questionnaire (GRWQ, ). SubjecSubjects attended 15 treatment sessions in groups of 32 to 35 women, for approximately 4 months. Average attendance to the treatment sessions was 83%. Sessions lasted 120 minutes and included educational content and practical application classroom exercises in the areas of physical activity and exercise, diet and eating behavior, and behavior modification . PhysicaMeasures of central tendency, distribution, and normality were examined for all psychosocial variables at baseline and for weight at baseline and 4 months. Following intention-to-treat principles and to include psychosocial data from all starting subjects in statistical analysis, the Last Observation Carried Forward (LOCF) method was used for 5 subjects who dropped from the program and could not be reached for testing at 4 months (the five subjects dropped after sessions number 10, 11, 12 [two subjects], and 14); in these cases, the last measured weight, which was assessed weekly for each woman with the same scale as used in laboratory testing, was entered as their final weight. The limitations of this method notwithstanding , variatiRank-order correlation (Spearman's ρ) was used to estimate the linear relationship between predictors and weight change. All but one among independent variables assessed at baseline displayed a non-normal distribution, warranting the use of this non-parametric technique. The dependent measure was expressed as the difference between baseline and 4-month weight. An alternative way to express weight results is to calculate the "residualized" value for 4-month weight, after the effect of baseline weight is removed (i.e., regressed out in linear regression). This method protects against overcorrection of the post by the pre score when using a subtraction score, and also effectively and completely adjusts this new "change" score for the pretreatment weight value . This vaQuadratic terms were produced for the two weight outcome evaluation variables, to assess the curvilinear relationship between these measures and actual weight results. Multiple regression analysis was performed to assess the multivariate relationships between the independent variables and weight change. In this regression model, the selected predictors  were forced into the model and the squared semi-partial correlation coefficient was calculated to quantify the unique contribution of each predictor to the variance in the dependent measure . ConsideWeight loss data reported in the present study refer to the initial 4 months of a longer trial. After the 4-month phase, subjects were randomly assigned to three distinct long-term interventions. Figure p = 0.480, comparing subjects responding "yes" and "no"). Two additional variables from the GRWQ were also analyzed. "Dream" weight (mean ± SD, 98.1 ± 3.9%) was unrelated to baseline-adjusted weight loss (ρ = 0.001, p = 0.98) while "disappointing" weight outcome  was associated with baseline-adjusted weight loss (ρ = 0.27, p = 0.002). Time at current weight, obesity-specific quality of life, self-esteem, and exercise variables were not associated with weight results, before or after adjusting for baseline weight. Significant predictors in the bivariate analysis .Weight outcome evaluations were computed as a percentage of participants' initial weight. Thus, the lower this percentage, the more stringent (i.e., more demanding) was a subject's evaluation of her results, and vice-versa. We found significant and positive linear relationships between outcome evaluations and weight loss Tables  and 2, ip = 0.284) and "acceptable" (p = 0.145) weight loss evaluations. Body size dissatisfaction scores were also not different among the three groups (p = 0.432). Table To further explore the association of the selected predictors with weight outcomes, subjects were divided into three groups based on tertiles of weight reduction adjusted for initial weight, and baseline psychosocial measures were compared among groups. Significant overall (ANOVA) differences emerged for the number of previous diets and self-motivation, with post-hoc comparisons showing significant mean differences only between the most and least successful groups Figure . Consides p = 0.42. Table This study aimed at reexamining the association between several pretreatment individual characteristics and success in short-term behavioral weight reduction, in overweight and moderately obese women. Ten variables which had previously been shown to predict weight change  were anamechanisms (mediating variables) by which treatments affect subjects, and for whom treatments are most effective . The potential benefits of studying moderators and mediators of outcomes within the behavioral and social sciences, including for physical activity, diet, and weight control are well described in the literature [To date, the majority of research on the treatment of overweight and obesity has focused on assessing overall treatment efficacy  and analyzing which programs work best, typically using randomly-assigned experimental treatment groups -27. By cterature -30. The terature .diets have you started...?", could also be a marker for more extreme dieting behaviors that may not be sustainable after the initial boost of motivation [In the present study and in other trials -36, prevtivation . This coFour earlier reports have examined the role of self-motivation as a predictor of weight loss ,36,40,41This study is among only a few that have analyzed associations between the Goals and Relative Weights Questionnaire and subsequent weight loss. Interestingly, marked differences emerged between the present and two previous analyses ,36. Portand moderate expectations/outcome evaluations foretell the best overall results, particularly if accompanied by a high sense of self-assurance [In the present study, it appeared that the weights participants would find acceptable/happy were associated with weight loss  until a certain threshold was reached, somewhere around 85 to 90% of initial weight (10–15% weight loss); for women reporting outcome evaluations below that level no further benefit was apparent. One previous study has shown that women with more modest absolute weight loss goals were more likely to achieve their goals, and that those who achieved their weight goals had better weight maintenance after 2.5 years; however, desired weight loss did not directly predict actual weight loss . Positivssurance .how much weight do you think you will lose by the end of this program?), while simultaneously tapping into a more attitudinal facet towards a person's weight and weight loss (how happy/accepting/disappointed would you feel at certain levels of weight loss?). To some extent, the latter could measure idealization of body weight and perceived importance of body weight and shape for self-esteem and well-being. Therefore, it is possible that moderate or "realistic" weight outcome evaluations  are the most beneficial and indeed reflect a good balance between a sufficient and necessary sense of self-efficacy and low to moderate levels of thin-ideal internalization, a variable which has been shown to be a positive risk factor for body dissatisfaction, negative affect, and eating disorders [It should be noted that variables originating from the GRWQ are closely related but are not equivalent to the construct of outcome expectancies (the belief that certain actions will lead to the projected results ) or to wisorders ,56.p = 0.042), the number of previous diets (ρ = 0.22, p = 0.013), and weight-related quality of life (ρ = -0.37, p < 0.001). Rapid and concurrent improvements in body image and eating behavior (e.g., reduction in binge episodes) have been observed after surgery-induced thinning [Women reporting a larger discrepancy between self and ideal body figures, which indicates a higher body size dissatisfaction , were lethinning , clearlythinning . Despitethinning  and the thinning , differethinning ,58.Strengths of this study are the a priori selection of variables to be analyzed as predictors, a unique population (Portuguese women), and the very low dropout rate. Limitations include a moderately-sized sample considering the known measurement error associated with questionnaire psychological assessments, the fact that some of the scales used still lack well-established validity, and the absence of a control or comparison group.Several pretreatment variables were re-evaluated as predictors of short-term weight loss in women. Previous dieting, low self-motivation, and body size dissatisfaction were confirmed as negative predictors of weight outcomes, while the relationship of outcome evaluations with weight reduction suggested a negative and curvilinear pattern, with positive but not excessively demanding evaluations presaging the best results. These data regarding people's outcome evaluations prior to weight loss may have important clinical implications  and are None declared.PJT conceived the study, led the implementation team, performed most statistical analysis, and drafted the manuscript. ALP participated in the study's implementation, in statistical analysis, and was responsible for all psychometric assessments. TLB, SSM, CSM, and JTB were actively involved in the study's implementation and in data collection. AMS participated in the study's implementation and collected all body habitus data. LBS is the principal investigator in the research trial and contributed to the final version of the manuscript. All authors read and approved the final manuscript.