, 2012) lacks supporting evidence. Human skeletons in the Peruvian Amazon, Santarem area, and middle Orinoco show little or no isotopic effect of maize until late prehistory ( Roosevelt, 1989, Roosevelt, 1997 and Roosevelt, 2000:482–485), when open-field maize cultivation is recorded in floodplains

and wetlands. The sun-loving grass maize (Zea mays, Poaceae) was an introduced cultigen (no wild relatives are known for South America), selleck whereas most Native Amazonian cultigens tend to be grown in mixed slash and burn fields, like manioc (Manihot esculenta, Euphorbiaceae) ( Olsen and Schaal, 1999), or in mixed orchards of the domesticated peach palm (Bactris gasipaes) and fruit trees that, though not domesticated, were cultivated ( Clement, 1999, Clement et al., 2010, Mora-Urpi et al., 1997 and Smith et al., 2007). Although Amazonia’s most important crop plant was the shrub Selleck JNJ26481585 manioc, the second most important domesticate original to Amazonia was the peach palm, and the majority of other plants cultivated by Amazonians are woody trees ( Clement et al., 2010:74). Prehistoric earthworks are another important human alteration to Amazon landscapes (Roosevelt et al., 2012 and Roosevelt, 2014). Amazonian mounds were built to elevate surfaces for residential, social, ritual, symbolic, defensive, transportation,

or agricultural purposes. Some raised settlements

above flood level, creating ponds with their borrow pits. Some seem to make sociopolitical or religious statements: to raise some residences above others, to bring cemeteries into more prominence, or to create ritual precincts and shrines. Transportation structures range from for causeways to ritual promenades and channels for boats. Agricultural works range from raised field surfaces to drainage ditches. While residential mounds are packed with rich, dark refuse, other structures, facilities, and especially socio-technic constructions can be almost devoid of refuse except for rare, cached offerings. Platform mounds for structures also can be almost devoid of artifacts except for their upper surfaces, as can raised fields. But all these structures include some kind of macroscopic or microscopic specimens and chemical and sedimentological evidence of their origins and use as human artifacts. One of the earliest and largest examples of extensive terra firme earthwork systems are those of the Faldas de Sangay culture of Ecuador in the western Amazon ( Porras, 1987, Rostain, 2010, Rostain, 2012, Salazar, 1998 and Salazar, 2008). Lying below the recently extinct volcano Sangay, it is a hilly tropical forest area drained by the Napo and its tributaries. Most of the current surfaces are quite rich tropical soils derived from the weathering of volcanic rocks and ash.

2 Gy of EBRT. Biochemical control was observed in 88% of patients with a median followup of 30 months (19). Burri et al. have reported the outcomes of 37 patients with a median followup of 87 months who underwent low-dose-rate salvage brachytherapy.

At 5-year followup, 65% of patients were found to have achieved biochemical Etoposide purchase freedom from failure (Phoenix definition), and 57% remained free of biochemical failure at 10 years. Grade 3 or 4 toxicity was noted in 11% of patients, consisting of transurethral resection of the prostate (TURP) in 2 patients, hematuria requiring fulguration in one patient, and one case of prostatorectal fistula (20). Several aspects unique to HDR brachytherapy make it ideally suited for use as a salvage procedure. • Brachytherapy delivers high-dose radiation directly to the target tissue. In the case of EBRT, radiation must pass through skin, muscle, bone, bowel, and other soft tissues to reach the prostate. Thus, in the setting of previously irradiated patients, HDR brachytherapy can deliver radiation without repeating significant exposure to these surrounding tissues. In addition, a patient-based dosimetric comparison between stereotactic body radiotherapy and HDR brachytherapy found that EBRT was not able

to achieve either the high doses to the prostate or the dose-sparing effect on normal tissues that HDR brachytherapy selleck kinase inhibitor is able to achieve (21), underscoring the advantages of HDR over external beam techniques. The results of the University of California San Francisco (UCSF)

salvage HDR experience have been recently updated (7). In this retrospective analysis, 52 consecutive patients received 36 Gy in six fractions, given in two insertions, 1 week apart. With a median followup of 60 months, 51% of patients were found to be biochemically free of relapse at 2 years, and only two patients developed Grade 3 or higher GU toxicity using CTAE criteria. An initial report published almost in 2007 reported 14% Grade 3 complications in 21 patients with a median followup of 19 months (22), suggesting that, with further time, Grade 3 complications tend to improve. The results of the USCF experience as well as the current report have used dose-fractionation schedules with similar biologic effective doses (170–180 Gy, assuming an α/β of 1.5 Gy) with acceptable toxicity and tumor control, suggesting that either 36 Gy in six fractions with two insertions, or 32 Gy in four fractions in a single insertion, would be appropriate fractionation schedules to consider. However, the optimal dose-fractionation schedule for salvage HDR brachytherapy has yet to be elucidated. As illustrated in Table 3, the outcomes of the present study compare favorably with results in the literature. HDR brachytherapy is an excellent treatment for salvage because it provides the high doses for hypofractionation while maintaining low doses to the urethra, bladder, and rectum.

ZR2010CM044), the National Basic Research Program of China (973 Program, Grant No. 2009CB118602), and State Key Laboratory of Crop Biology (Grant No. 2012KF01) of Shandong Agricultural University, Tai’an, Shandong, China. “
“Rice blast disease, caused by the filamentous ascomycete fungus Magnaporthe oryzae (formerly Magnaporthe grisea), is one of the most destructive diseases of rice worldwide. The fungus can also cause severe infections in wheat [1]. Avirulence (AVR) genes in M. SB431542 solubility dmso oryzae are known to be highly unstable [2] owing to

the presence of a large number of active transposable elements that can shape the evolution and adaptation of the fungus [3]. To date, seven race-specific avirulence genes, AVR-Pita1, AVR1-CO39, ACE1, AVR-Pizt, AVR-Pik, AVR-Pii and AVR-Pia, have been molecularly characterized. ACE1 has been found among rice isolates and can be used to induce avirulence [4]. It encodes a putative polyketide synthase fused to a non-ribosomal peptide synthetase determining the specificity of Pi33 [4]. AVR1-CO39, an isolate of M. grisea from weeping lovegrass, encodes a signal that triggers a strong defense response in the rice cultivar CO39, which www.selleckchem.com/screening/anti-diabetic-compound-library.html carries the corresponding resistance (R) gene [5]. AVR-Pizt, recognized by the R gene Piz-t, can suppress BAX-mediated

programmed cell death in Nicotiana benthamiana, suggesting a mechanism by which AVR-Pizt may confer virulence on M. oryzae [6]. The remaining three Edoxaban AVR genes of M. oryzae were novel

genes [7]. In contrast, abundant major and minor blast resistance (R) genes have been identified in rice germplasm worldwide [8]. Thus far 16 major and two minor blast R genes have been cloned, most of which encode NBS type R proteins [9]. Understanding R–AVR gene interaction specificity is important for the development of effective strategies to manage rice blast disease. AVR-Pita determines the efficacy of the R gene Pi-ta [10]. The genes Pi-ta and AVR-Pita are the first R/AVR gene pair characterized in the rice blast system. AVR-Pita is located in the telomeric region of chromosome three of M. oryzae, and was cloned from a Chinese isolate, O-137 [10]. AVR-Pita was renamed AVR-Pita1 following the discovery that it has several family members in the Magnaporthe species [11]. AVR-Pita1 encodes a protein with 223 amino acids with properties highly similar to those of a metalloprotease [10]. AVR-Pita716 codes for a putative product that was predicted to be an elicitor that binds to Pi-ta directly inside plant cells, triggering an effective blast resistance response [12] and [13]. Other biological functions of AVR-Pita in M. oryzae remain unclear; however, it was predicted to be involved in the early stages of pathogenesis [10]. Recently, AVR-Pita1 was shown to accumulate in the biotrophic interfacial complex (BIC), raising the possibility that AVR-Pita1 prepares rice cells for subsequent fungal entry and biotrophic growth [14]. A recent analysis of strains of M.

, 2009). Importantly, these structural analyses indicate that antigen recognition by VLR antibodies is distinct from antigen recognition by conventional immunoglobulin-based antibodies. The unique origins and structural characteristics of VLR antibodies suggest that these proteins have the potential to complement conventional antibodies in biomedical

research applications and for biomarker check details discovery studies. Here we describe the generation of monoclonal VLR antibodies to human T lineage lymphocytes and demonstrate applicability of monoclonal VLR antibodies for affinity purification and mass spectrometric identification of the cell surface antigens. Lamprey larvae (80–100 mm, Lamprey Services, Ludington, MI) in length were anesthetized (0.1 g/l MS222/0.14 g/l sodium bicarbonate) and immunized with 2 × 106 www.selleckchem.com/products/cx-5461.html primary lymphocytes enriched for CD4+ T cells in 60 μl of 0.66 × PBS. The animals were boosted twice at 2 week intervals with an equal number of cells obtained from different donors to avoid the generation of alloantigen-specific VLRs. 10 days after the second boost the animals were sacrificed (1 g/l MS222/1.4 g/l

sodium bicarbonate) followed by exsanguination. Peripheral blood was collected in 0.66 × PBS/30 mM EDTA, layered on top of 55% percoll and subjected to density centrifugation (400 ×g, 20 min). Subsequently, the lamprey lymphocytes were collected and the antisera were analyzed for reactivity to primary human PBMC. Out of 3 immunized animals, we chose the animal with the highest polyclonal VLR antibody new titer for subsequent expression library generation. Peripheral blood was obtained from healthy volunteers of the Vaccine Center of Emory University, Atlanta, GA after informed consent was obtained. Tonsil samples were obtained from Children’s Healthcare of Atlanta and chronic lymphocytic leukemia (CLL) samples from Emory University tissue procurement facility. All studies with human tissues were approved by the Institutional Ethics Review Board and were conducted in accordance

with institutional guidelines and the declaration of Helsinki. Tonsilar single cell suspensions were generated by tissue mincing, filtration through 70 μm wire mesh, and cell centrifugation on a ficoll-hypaque gradient. Blood CD4+ T cells were purified using CD4 microbeads (Miltenyi Biotec, Cambridge, MA) followed by magnetic separation. Hemopoietic cell lines were maintained in RPMI 1640 supplemented with 10% fetal calf serum, 2 mM l-glutamine, 100 U/ml penicillin/streptomycin, and 50 μM β-mercaptoethanol and HEK293T cells were maintained in DMEM supplemented with 10% fetal calf serum, 2 mM l-glutamine, and 100 U/ml penicillin/streptomycin. Antibodies to CD3, CD5 and CD19 were obtained from BD-Biosciences (San Jose, CA).

Each image was transferred AZD2281 ic50 to a SIS AnalySIS FIVE database (Soft Imaging System GmbH, Münster, Germany). In addition, all data of the image analysis were entered into the program and stored in this database. The methods and results of the histopathological examination have been published elsewhere (Ernst et al., 2002, Ernst et al., 2005 and Kolling et al., 2008). The inflammation score, which was correlated with the genotoxicity markers, was based on a grading scheme comprising five degrees of alterations (0 = none; 1 = very slight; 2 = slight; 3 = moderate;

4 = severe; 5 = very severe). Grade 1 (very slight): <10% of lung tissue affected; For statistical analysis, the SAS software package (release 9.1 on Windows XP computer, SAS Institute, Cary, NC, USA) and Statistica (version 8.0, StatSoft Inc., Tulsa, OK, USA) were used. The image analysis data were imported into these software packages. find more Data were analyzed by using analysis of variance (ANOVA) as an overall test. If the group means differed

significantly by ANOVA, the treatment groups were compared with the control group using Dunnett’s test. The Tukey HSD test was used as another post hoc test for comparison among the different treatment groups, as this test is not based on comparison between treatment and control. Statistical significance was reached if p ≤ 0.05. Data were considered highly significant if p ≤ 0.01 or p ≤ 0.001. The data for evaluation of possible correlations between genotoxicity marker expression and, for example, histopathology or BAL data were extracted from the interim and final reports of the research projects of the German Federal Environment Agency (Ernst et al., 2002, Ernst et al., 2005, Kolling et al., 2008 and Kolling et al., 2011) and from the raw data of individual animals of the research projects. Correlations between the genotoxicity markers and other study parameters such as inflammation score and enzymatic activities or cell acetylcholine counts in BAL fluid were calculated

using the respective group mean values. In case of the inflammation score, the individual animal data were additionally used for determination of correlations, because the same animals were investigated for both genotoxicity marker expression and histopathologic evaluation of lung inflammation. The method of linear regression/Pearson product-moment correlation (SAS [Cary, NC, USA] software package Statistica or SigmaStat 3.1) was used to calculate the correlation coefficient (r) and the significance of the correlation (p-value). Correlation coefficients lacking statistical significance were rated as “correlation without significance” if r > 0.5. PAR synthesis was determined in particle-exposed lung tissue as a general marker for genotoxic stress. Three months after the first and one month after the last exposure, alveolar lining cells of quartz DQ12-treated rats showed a statistically significant, about 1.

To get adequate coverage on the left seminal vesicle and left base, 1.2 cc of the bladder was allowed to receive 75% of the prescription dose, slightly exceeding the 1 cc goal. However, there was a dramatic dosimetric decrease in the rectum owing to the spacer. Although the goal was to

keep less than 1 cc of the rectum to 75% of the dose, there were 0 cc of rectum receiving 75% of the dose, as seen in Fig. 3, where the 75% isodose line is entirely within the spacer and does not touch the rectum. A small amount of the rectum was within the 50% isodose line, and the radiation dose to the hottest 2 cc of the rectum was approximately 3 Gy per fraction. The patient had no urinary frequency, nocturia, or hematuria. Nine months after implant, the patient developed

mild rectal bleeding which was eventually treated with argon plasma coagulation at month 12. Ganetespib clinical trial For men who develop prostate cancer after prior pelvic radiotherapy, the available treatment options are limited (3). Most of the world’s literature on the subject is from men who received prior radiation for prostate cancer (typically to a dose of approximately 70 Gy) and then recurred. Major approaches that have been attempted with curative intent include radical prostatectomy, brachytherapy, and cryotherapy. Performing a salvage radical prostatectomy PD-0332991 mw in a radiated field can be difficult and lead to high complication rates. Series have reported up to a 67% rate of some degree of incontinence (4), 15% rate of rectal injury (5), and 29% rate of bladder neck stricture (4). Of all 531 cases of salvage prostatectomy that had been published

in the English literature from 1990 to 2007, the rate of incontinence Pyruvate dehydrogenase was 41%, rectal injury was 4.7%, and bladder neck stricture was 24% (3). Cryotherapy is not widely used as a first-line option for the definitive treatment of prostate cancer, and it is unknown whether its efficacy would be similar to surgery or radiotherapy. In the postradiation setting, cryotherapy has been associated with up to a 96% rate of incontinence (6), a 55% rate of urethral sloughing (6), 55% rate of bladder stricture (7), 44% rate of perineal pain [8], [9] and [10], and 11% rate of fistula (7). Of the 510 cases of salvage cryotherapy reported from 1990 to 2007, a weighted average of morbidity yields a 36% rate of incontinence, a 11% rate of urethral sloughing, 17% rate of bladder stricture, 36% rate of perineal pain, and 2.6% rate of fistula (3). Salvage brachytherapy after prior radiotherapy has also been reported, either as low-dose-rate seed implantation or HDR implantation of empty catheters into which a highly active radioactive source is placed for precise amounts of time to create the appropriate dose distribution.

Some of these recurrent CNVs coincide with known

genomic disorders, whereas others involve genes associated with ASD or developmental delay and intellectual disability [ 28 and 30]. In studies of idiopathic ASD, the most common recurrent anomaly is a ≈600 kb microdeletion/microduplication of chromosome 16p11.2 (∼0.8%) [ 22, 44• and 45]. This CNV is also observed in ASD cases with additional dysmorphology [ 46 and 47], in non-ASD cases with developmental delay [ 48, 49 and 50] and/or obesity [ 51, 52 and 53], in subjects with various non-ASD psychiatric disorders [ 44•, 54, 55, 56 and 57], and in some apparently unaffected individuals [ 48]. The 16p11.2 deletions appear learn more more penetrant (nearing 100% for either ASD or developmental delay)

than the duplications (∼50% penetrance); (vi) there is enrichment for gene-rich CNVs, and especially CNVs that comprise neuronal synaptic complex genes [ 58, 59 and 60]. Finally, (vii), while a number of CNVs appear to involve haploinsufficient regions, or to act dominantly, others appear to act recessively, such as PCDH10, DIA1, and NHE926 – identified by a study of consanguineous INCB018424 concentration ASD families and rare homozygous CNVs that deleted both copies of these genes [ 61]. It is probable that different CNVs exhibit different penetrance for ASD depending on the dosage sensitivity and function of the gene(s) they affect [28 and 60]. Some CNVs have a large impact on ASD expression (e.g., 15q11–q13 duplication); these will typically be de novo in origin, cause more severe ASD symptoms, and be more prevalent among sporadic ASD. Other CNVs have moderate or mild effects (e.g., 15q11.2 deletion) that probably require other genetic (or non-genetic) factors to take the phenotype across the ASD threshold. Some of these CNVs demonstrate variable phenotypic expression, are found in other disorders,

or are observed in non-ASD relatives and some population controls. CNV screening Phospholipase D1 and direct sequencing of candidate genes are rapidly identifying genes for further characterization in relation to ASD. These approaches have implicated NLGN3 [ 62••], NLGN4 [ 62•• and 63], SHANK2 [ 20••, 64, 65 and 66], SHANK3 [ 67•• and 68], NRXN1 [ 31••, 69 and 70] and NRXN3 [ 71], PTCHD1/PTCHD1AS [ 20•• and 72•], SHANK1 [ 73], DPYD [ 24 and 74], ASTN2 [ 34 and 57], DPP6 [ 22], MBD5 [ 75, 76 and 77], CDH8 [ 78] and CNTNAP2 [ 79] (among others) as affecting ASD risk. Some rare, highly penetrant mutations appear as sufficient to be monogenic causes of ASD ( Figure 2). At this writing, large-scale sequencing projects have been initiated, to target the majority of genic regions (or exome) from hundreds of families with ASD. Three studies [80, 81 and 82], which studied over 600 ASD families, report on de novo variants in these families. All find a two-fold to four-fold increase in de novo nonsense variants among affected subjects over that expected by chance.

The favoring of a pseudoplastic behavior probably occurred due to higher-molecular-weight polymers formed during the cross-linking reaction promoted by the TG. Innocente, Comparin, and Corradini (2002) affirmed that with an increase in the shear rate, large polymer molecules tend to disentangle and possibly align in the flow field, offering less resistance to flow. The rheological behavior of the samples after the reduction of shear rate (downward curves) can

be seen in Table 3. Using the Power Law model it was observed that K varied from 0.08 to 0.15 Pa s−1, values Androgen Receptor Antagonist which are lower than those obtained for the upward curve. All samples containing TG had higher K values than the respective samples without TG, demonstrating that the addition of TG gives the ice cream a greater resistance to structural rupture. Moreover, the values of the flow behavior index (n) were greater than those of the upward curve, showing that there was a decrease in the pseudoplastic properties when the shear rate decreased.

The decrease in K and increase in n can be attributed to the structural breakdown Erlotinib cost of the protein network of the ice cream due to shearing, which favors this behavior. An important feature of the shear stress versus shear rate results, obtained by increasing and then decreasing the shear rate, is the formation of Methocarbamol hysteresis. The area formed between the curves indicates that the fluid viscosity is time dependent (Tárrega, Durán, & Costell, 2004). Table 4 shows the hysteresis values for the ice cream samples. It can be observed that the TG addition caused an increase in the degree of hysteresis when compared with the controls

(without TG). Samples IC4-TG and IC6-TG (Table 4) showed the greatest degree of hysteresis with no significant differences (P < 0.05) between them. This demonstrates that these two samples needed more energy to break the ice cream structure formed from the protein polymerization, providing a firmer product. IC4-TG and IC6-TG were also the samples that showed the highest apparent viscosity and consistency index. According to Tárrega et al. (2004), a high-viscosity thixotropic fluid may show a larger hysteresis area than a lower viscosity one, even if the latter undergoes a more accentuated destruction of the structure. The presence of hysteresis was also observed by González-Thomás et al., (2008) and Karaca et al. (2009) in studies on ice cream. The time-dependent rheological data were fitted using the Weltman model in order to characterize the thixotropic behavior of the ice cream samples. It was observed that the TG addition resulted in a significant increase in the initial tension required (A) to initiate the breaking of the ice cream structure ( Table 5) due to the formation of a more stable network.

For these reasons, we reject the view the NMAs merely represent unnatural disruption of actions caused by stimulating areas normally involved in positive movement generation. An alternative possibility remains open: negative motor responses might represent an artificial induction of a normal physiological process of action inhibition. In our view, the normal organization of complex (Gerloff et al., 1997) and fine movement (Fukaya et al., 2004) involves an element of inhibition. Hierarchical control is required to regulate the balance of activation and inhibition in several motor cortical areas, so that movements are neither hyperkinetic and impulsive, nor hypokinetic and ineffective.

Crucially, we suggest that there is some ‘functional truth’ in NMAs. We speculate that DES, albeit not ecological itself, produces negative motor responses by activating physiologically

inhibitory pathways that participate in selleck normal action control. Crucially, negative motor responses are not simply an artifactual, unnatural disruption of ongoing movement, or an overloading of positive motor effects. The interesting observations reported by Swann et al. (2012) provide clear, and perhaps the first, evidence for a possible functional relevance of NMAs in action inhibition, as an important element of action control. The natural inhibitory function of NMAs could be important in action control for two distinct reasons. First, NMAs may reflect activation of an inhibitory mechanism for praxic control of fine details of action execution. Dapagliflozin mw Alternatively, NMAs may reflect artificial activation of an inhibitory mechanism for executive, decisional

control over whether actions occur or not. The data reviewed here cannot conclusively distinguish between these two alternatives, and future functional studies may shed light on this interesting question. Control of praxis has been strongly linked to lateral cortical pathways linking the inferior parietal cortex and the lateral premotor cortex (Tanji and Hoshi, 2008). In contrast, executive control of action has been linked to the prefrontal and medial frontal cortices (Badre and D’Esposito, 2009 and Stuss and Knight, 2002), and particular to the drive these areas receive from the basal ganglia (Heyder et al., MRIP 2003). Our review shows two clear clusters of NMAs in the lateral frontal and dorsomedian frontal cortices. By analogy with the lateral/frontal division for positive motor function, we can thus speculate that the lateral frontal cluster of NMAs reflects a praxic mechanism for fine regulation of complex action sequences, while the medial frontal cluster represents an executive mechanism for regulating whether an action is executed or inhibited. From the evidence reviewed above, we suggest that NMAs are indeed truly inhibitory.

The random null model was of equal proportions of positive, neutral and negative effects, while the no-effect null model was that coinfecting pathogens do not interact,

allowing for a 5% error rate (hence 2.5% negative, 2.5% positive, and 95% neutral reported effects). PI3K inhibitor drugs This constitutes a recommended vote-counting method deriving continuous parameters analysed against confidence intervals (α = 0.05). 27 Finally, we explored the potential influence of the missing data (NAs) on the effects of coinfection in the analysis (56 for pathogen abundance, 79 for host health). These values represent reported coinfections where the effect on either pathogen abundance or host health was not reported, despite the possibility that these coinfecting pathogens did interact with each

NU7441 other and/or influence host health. We therefore assessed how potential interactions from these unreported effects may alter the overall patterns of coinfection effects. To determine their potential impact on the estimated overall effects, NAs were assigned one of three values at random (+1, 0, −1). The mean effect was then calculated per publication or coinfection pair as before, and a grand mean taken across all publications or coinfection-pairs. The grand mean represents an estimate of overall effect of coinfection on either host health or pathogen abundance across either publications or coinfections, given a particular random assignment of −1, 0, +1 to NAs. Repeating this random assignment 1000 times produced a distribution of grand means. We examined whether recent coinfection research focuses on the pathogens causing the highest global mortality. We obtained global totals for the number of deaths (both sexes, all ages) in 2009 under every category of infection collated by the World Health Organisation

(obtained from the Global Burden of Disease section of the Global Health Observatory website).28 We compared the ten categories causing most global deaths in 2009 with total reports of coinfection involving these infections. Comparing the top ten infection categories by mortality with their morbidity measures (DALYs) yielded Tau-protein kinase similar trends, so we present only data from the mortality comparison. Hundreds of publications on coinfection are published annually and have increased from 219 publications in the first year of search results to 1464 publications in 2009 (Fig. 1). This increase includes studies of both human and non-human hosts. Of the 1464 publications retrieved for 2009, 309 reported multiple pathogen species coinfecting humans. Publications came from 192 journals, with most (136 of 192 journals, 70.8%) publishing a single coinfection article in 2009. The majority of relevant publications from 2009 were observational studies (234 of 309, 75.0%), of which 159 (67.9%) involved patient groups, 60 (25.6%) were case notes and 18 (7.7%) surveyed a population. Three observational studies (1.