Ultrasound-guided fine-needle aspiration cytology significantly improved mycetoma diagnosis.

BACKGROUND: Ultrasound (US)-guided fine-needle aspiration cytology (US-FNAC) has improved the diagnosis of many malignancies, infections and other diseases as it is safe, simple, quick and accurate. In mycetoma, it is assumed that this technique may have a better diagnostic yield than the conventional FNAC as it can accurately identify the optimal site for the aspiration. OBJECTIVE: To compare the diagnostic yield of conventional FNAC with US-FNAC. METHODS: This descriptive cross-sectional hospital-based study included 80 patients with clinically suspected mycetoma. RESULTS: Of the 80 patients included, 35 proved to have actinomycetoma, and 37 had eumycetoma based on surgical biopsies, histopathological examination and the culture of grains. Eight patients appeared to have no mycetoma. For actinomycetoma diagnosis, the US-guided FNAC improved sensitivity to 97% and negative predictive value (NPV) to 83% compared to the conventional FNAC, which had 63% sensitivity; and NPV of 28%. No improvement was found for specificity. For eumycetoma, the conventional FNAC had 86.5% sensitivity, 100% specificity, 100% PPV and 37.5% NPV. The US-FNAC for the diagnosis of eumycetoma had 100% sensitivity and specificity. CONCLUSIONS AND RELEVANCE: The obtained results showed that US-FNAC is better than the conventional FNAC with lower false-negative results. It can accurately distinguish between the two types of mycetoma, allowing rapid initiation of proper treatment. The technique can be used in rural areas with low resources and for epidemiological surveys as a quick screening tool for patients suspected of mycetoma.

Value of Serum Adenosine Deaminase (ADA) in Distinguishing between Tuberculous and Non-tuberculous Lymphadenopathies.

Persistent lymphadenopathy with or without fever is often a diagnostic challenge to the physician which are usually caused by infection like tuberculosis, hematological malignancy (lymphoma, leukemia), connective tissue diseases (SLE, RA, Sjogren's syndrome etc.), sarcoidosis, storage diseases, drugs (like phenytoin) in Bangladesh. To establish the cause of lymphadenopathy, we need to do a good number of investigations including invasive tests like FNAC or histopathology of the involved lymph node. In many instances these are not possible due to unavailability or cost. But for last few years the adenosine deaminase is an enzyme involved in purine catabolism and its significance in the diagnosis of tuberculosis has been demonstrated by many studies. In addition to tuberculosis, elevated serum adenosine deaminase has also been found in lymphoma, sarcoidosis and some connective tissue diseases. The study was intended to assess if there are any significant diagnostic difference in the level of elevated adenosine deaminase between tubercular and different types of non tubercular lymphadenopathy. It included 68 patients, equally divided into two groups, tuberculous lymphadenitis and non-tuberculous lymphadenopathy. Epitheloid granuloma with caseation necrosis in biopsy or FNAC was taken as case definition of tuberculous lymphadenitis. Causes of non-tuberculous lymphadenopathy were established on the basis of clinical findings, laboratory investigations and histopathological diagnosis of biopsy or FNAc materials. This cross-sectional observational study was done in Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh over a period of one year and participants of 18 years and above of both genders were included as per consecutive sampling technique. Serum ADA concentrations were estimated by enzymatic method. Mean serum ADA concentration was 25.52±7.11 in tuberculous lymphadenitis and in non-tuberculous lymphadenopathy patients it was 27.29±15.91U/L with no significant difference (p=0.480). The non-tuberculous lymphadenopathy group consisted of Hodgkin disease (n=9), non-Hodgkin lymphoma (n=10), sarcoidosis (n=2), reactive lymphadenitis (n=9) and other lymphadenopathy group (n=4) (that consisted one case of each of follicular hyperplasia, adult Still disease, sinus histiocytosis and Castleman's disease). The mean ADA of these groups was 32.77±13.14U/L, 46.40±46.10U/L, 13.94±2.81U/L and 21.75±3.17U/L respectively. Tuberculous lymphadenitis patients had significantly higher serum ADA than persistent reactive lymphadenitis. On the other hand, there were statistically significant elevation of serum ADA in non-Hodgkin lymphoma and sarcoidosis than in tuberculous lymphadenitis.

Nano BEA zeolite catalysts for the selective catalytic cracking of n-dodecane to light olefins.

Nano BEA zeolite catalysts were synthesized and modified by desilication and then ion-exchanged with Co. The desilication was carried out using 0.1 M of NaOH. The synthesized and modified nano BEA catalysts were characterized via different characterization techniques. Ammonia temperature program desorption (NH3-TPD) and the pyridine Fourier transform infrared (pyridine-FTIR) were utilized to investigate the acidity of catalysts. X-ray diffraction (XRD), 27Al and 29Si nuclear magnetic resonance (NMR) spectroscopy techniques were used to examine the structure of the catalysts. The XRD patterns of the as-synthesized nano BEA catalysts were identical to that of the reference, while the NMR analysis revealed the distribution of silicon and aluminum in the BEA structure. The scanning electron microscope (SEM) analysis confirmed that the fabricated catalysts were less than 100 nm. The desilication and Co ion-exchange altered the acidity of the catalyst. The catalysts were evaluated in the cracking of sssssss to light olefins in the temperature range from 400 °C to 600 °C. The conversion increased with the increase in the reaction temperature for both catalysts; the conversion was above 90% for the Co-BEA catalyst at a temperature above 450 °C. The yield of light olefins also increased at higher temperatures for both catalysts, while at a lower temperature the yield to light olefins was ca. 40% over that of Co-BEA.

HIV-1 infection of CD4 T cells impairs antigen-specific B cell function.

Failures to produce neutralizing antibodies upon HIV-1 infection result in part from B-cell dysfunction due to unspecific B-cell activation. How HIV-1 affects antigen-specific B-cell functions remains elusive. Using an adoptive transfer mouse model and ex vivo HIV infection of human tonsil tissue, we found that expression of the HIV-1 pathogenesis factor NEF in CD4 T cells undermines their helper function and impairs cognate B-cell functions including mounting of efficient specific IgG responses. NEF interfered with T cell help via a specific protein interaction motif that prevents polarized cytokine secretion at the T-cell-B-cell immune synapse. This interference reduced B-cell activation and proliferation and thus disrupted germinal center formation and affinity maturation. These results identify NEF as a key component for HIV-mediated dysfunction of antigen-specific B cells. Therapeutic targeting of the identified molecular surface in NEF will facilitate host control of HIV infection.

Assessment of genetic diversity and population genetic structure of Carthamus species and Iranian cultivar collection using developed SSR markers.

This experiment was conducted to assess genetic relationships among safflower genotypes from different geographical regions of Iran and other countries using newly developed simple sequence repeat (SSR) markers. By enrichment method, 32 primer pairs were designed of which 18 pairs were able to detect polymorphism in 105 safflower cultivars from Carthamus tinctorius, C. oxyacanthus, C. lanatus, C. glaucus, C. boissieri and C. dentatus. The selected SSR primers amplified a total of 59 alleles with an average of 3.27 alleles per locus among the cultivars and the average values of gene diversity, heterozygosity and PIC were 0.45, 0.37 and 0.39, respectively. Neighbour-joining cluster analysis based on Nei's genetic distance categorized populations of Carthamus in six major clusters; all wild accessions were grouped differently from cultivated genotypes. Cluster analysis significantly distinguished C. oxyacanthus genotypes in different categories: centre (Arak), northeast (Azarbaiejan), east (Kermanshah) and southeast (Shiraz, Chaharmahal and Kohgiluyeh). The presence of C. boissieri and C. glaucus in one cluster appeared to be in close relationship between each other, indicating a common ancestor. The results revealed that C. dentatus discriminated from the species with n = 10 chromosomes, C. boissieri and C. glaucus assigned in to separate subsection. In summary, this study has shown that domesticated and wild genotypes were clustered into two major groups indicating these markers as appropriate tools to amassment genetic diversity and genome mapping.

Probiotic feeding affects T cell populations in blood and lymphoid organs in chickens.

This study was performed to evaluate the effects of Lactobacillus acidophilus bacteria as a probiotic on chicken T cell subset populations in peripheral blood and lymphoid tissues. Thirty chickens were divided into three groups and fed sterilised cow milk, a mixture of milk and L. acidophilus (probiotic), or neither, as the control group. Chickens were euthanised after 14 and 21 days, and whole blood and ileal, bursal, and caecal tonsillar tissues were collected. The populations of T cell subsets, including CD4+, CD8+, and TCR1+ cells, were evaluated by immunohistochemistry and flow cytometry. After 21 days of treatment the percentage of blood CD4+, CD8+, and TCR1+ cells was significantly higher in the probiotic-fed group than in the control group. After 14 days of treatment, a significantly greater number of CD4+ T cells were found in the ileum of probiotic-fed chickens than in chickens from the other two groups. This difference was even greater after 21 days. In addition, after 21 days, a significantly greater number of TCR1+ cells were found in the caecal tonsils of milk-fed chickens than in chickens from the control group. The findings indicate that probiotics may alter the distribution of T cells in the blood and lymphoid tissues in young chickens; however, transient changes in lymphoid tissues indicate that probiotics likely do not permanently affect mucosal immunity.

Pepsin and bile acids in saliva in patients with laryngopharyngeal reflux - a prospective comparative study.

OBJECTIVES: Laryngopharyngeal reflux (LPR) and biliary duodenogastric reflux can cause damage to the laryngeal mucosa and voice disorders. The aim of this study was to find out whether levels of pepsin and bile acids in the saliva can serve as diagnostic markers of LPR. SETTING: A prospective comparative study. PARTICIPANTS: Twenty-eight patients with LPR proven via high-resolution manometry and combined multichannel intraluminal impedance and 24-h pH monitoring and 48 healthy controls without symptoms of LPR were included in the study. MAIN OUTCOME MEASURES: In the patients with LPR symptoms, oesophagogastroscopy with oesophageal biopsy was performed. The levels of total pepsin, active pepsin, bile acids and the pH of the saliva were determined in all participants and compared between the groups. Reflux symptom index (RSI) and reflux finding score (RFS) were also obtained and compared. The groups differed significantly in RSI (P = 0.00), RFS (P = 0.00), the levels of bile acids (P = 0.005) and total pepsin in saliva (P = 0.023). The levels of total pepsin and bile acids were about three times higher in the patients with LPR than in the healthy controls. There was a significant correlation between the RSI and RFS score and the level of total pepsin and bile acids in the saliva. Histopathological examination of the oesophageal biopsy taken 5 cm above the lower oesophageal sphincter confirmed reflux in almost 93% of patients with symptoms. CONCLUSIONS: The study results show that the levels of total pepsin and bile acids in saliva are significantly higher in patients with LPR than in the controls, thus suggesting this as a useful tool in the diagnosis of LPR and particularly biliary LPR.

New threshold temperatures for the development of a North American diamondback moth (Lepidoptera: Plutellidae) population and its larval parasitoid, Diadegma insulare (Hymenoptera: Ichneumonidae).

The currently accepted lower threshold temperature for the development of diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae), the world's most destructive insect pest of cruciferous crops, is around 6.0°C, and there is no known upper threshold temperature. Neither are there established threshold temperatures for diamondback moth's major natural enemy, Diadegma insulare (Hymenoptera: Ichneumonidae). Laboratory studies were undertaken to determine the survival and development of a North American diamondback moth population and its parasitoid D. insulare at 20 constant temperatures ranging from 2.0 to 38.0°C. Diamondback moth completed development from second instar to adult within a temperature range of 4.0-37°C, and D. insulare completed its life cycle from egg to adult within a temperature range of 4.0-33°C. The developmental data were fitted into one linear and four nonlinear models. Using goodness-of-fit and the ability to estimate parameters of biological significance as selection criteria, the Wang model was the most acceptable among the nonlinear models to describe the relationship between temperature and development of both species. According to this model, the lower and upper threshold temperatures for diamondback moth were 2.1 and 38.0°C, respectively, and for D. insulare they were 2.1 and 34.0°C, respectively. Based on the Degree Day model, diamondback moth required 143 d above the lower threshold of 4.23°C to complete the life cycle, while D. insulare required 286 d above the lower threshold of 2.57°C. This study suggests that temperatures during the crop-growing seasons in North America are not limiting factors for development of either diamondback moth or D. insulare.

The burn registry program in Iran - First report.

Burn injury is still a major problem in Iran, resulting in many reports which are usually dispersed, uncoordinated and probably unreliable. We created a burn registry comprised of a data entry program with 222 variables for each admitted burn patient. This program was established in August 2010 at the Iran University of Medical Sciences, Motahari Burn Hospital. We had 14,277 burn patients from August 2010 to August 2011, 877 of whom were admitted to the hospital. Of the patients, 65.9% were male and 34.1% were female. The age was 28.85 years (SD = 19.77). The most prevalent cause of burn was flame 78.5% (pipe propane gas 57.2; kerosene 19.9%). The mean total body surface area (TBSA) involvement was 23%. A total of 77.8% of patients were discharged with partial recovery, while mortality was 8.9%. The mean hospital stay was 14.63 days (SD =11.07). The program is designed to help understand the scope of burn injury in Iran, providing information on patients, etiology, and course of treatment. It also highlights differences between various parts of the country in terms of the causes and frequency of burn injuries. Moreover, the burn registry provides a basis for further research and surveys for treatment and preventive programs. Our results showed that, although Emergency Medical Services (EMS) staff are highly capable and well-trained, their coverage seems to be less than 50% and needs to be increased. Marriage status was shown to have no influence on the occurrence of burns, and among our patients, 57.0 % were poorly educated.

Les blessures par brûlure sont toujours un problème majeur en Iran, résultant dans de nombreux rapports qui habituellement sont dispersés, mal coordonnés et probablement peu fiables. Nous avons créé un programme de saisie de données de registre de brûlure avec 222 variables pour chaque patient brûlé admis. Ce programme a été créé en Août 2010 à l'Université des sciences médicales de l'Iran, à l'hôpital Motahari. A partir d'Août 2010 jusqu'à Août 2011 nous avons eu 14,277 patients brûlés, dont 877 ont été hospitalisés. Parmi les patients, 65,9 % étaient des hommes et 34,1% étaient des femmes. L'âge moyen était de 28,85 années (SD = 19,77). La cause de brûlure la plus courante était la flamme (en 78,5% des cas, dont 57,2 % étaiaent causés par le gaz tuyau de propane et 19,9% par le kérosène). La moyenne de la surface corporelle totale (SCT) brulée était de 23%. 77,8 % des patients ont reçu leur congé avec récupération partielle, tandis que la mortalité était de 8,9%. La durée moyenne d'hospitalisation était de 14,63 jours (SD = 11:07). Ce programme a été conçu pour aider à comprendre la portée des brûlures en Iran, fournissant des informations sur les patients, l'étiologie et le traitement. En outre, il met en évidence les différences entre les diverses régions du pays en termes de la fréquence et les causes des brûlures. En plus, le registre fournit une base pour des recherches et des enquêtes sur le traitement et la prévention des brûlures. Nos résultats ont montré que, bien que le personnel EMS sont très capables et bien formés, la couverture des EMS semble être inférieure à 50% et doit être augmentée. Selon les résultats, le statut de mariage n'a aucune influence sur la survenue de brûlures. Enfin, parmi nos patients, 57,0% étaient peu instruits.

Transient and permanent hypoparathyroidism following thyroidectomy.

AIM: This study was designed to identify the risk factors of transient and permanent hypoparathyroidism in patients undergoing thyroidectomy. We also studied the duration of transient hypoparathyroidism following thyroid surgery. METHODS: A prospective study was conducted on 163 consecutive patients undergoing thyroidectomy. Serum Levels of calcium and parathyroid hormone (PTH) were determined the day before surgery and then on the 1st and 10th postoperative day to determine the postoperative course of serum PTH and calcium. Patients were followed for six months after surgery. RESULTS: The incidence of transient and permanent hypoparathyroidism was 8.6% and 0%, respectively. Among all of variables, only the extent of surgery had a significant relationship with postoperative hypoparathyroidism (P=0.010). PTH measurement at 10th day postoperatively became normal, except in one patient in whom it continued to be low until two months. CONCLUSION: The extent of surgery is the leading cause of postoperative hypoparathyroidism, though, in most of the patients this is a transient complication and PTH level revert to normal within 10 days after surgery.

Efficacy of Microwave and Infrared Radiation in the Treatment of the Skin Lesions Caused by Leishmania major in an Animal Model.

BACKGROUND: Cutaneous leishmaniasis caused by L. major is an important public health problem in endemic areas. The aim of this study was to explore the therapeutic effect of microwave and or infrared radiation in the treatment of lesion induced in BALB/c mice by L. major inoculation. METHODS: The footpad lesion was induced in BALB/c mice by inoculation of L. major promastigotes subcutaneously. The lesion was treated with 600 watts power, 2.450 GHz frequency and/or infrared device with 150 watts and a wave length of 890 nanometres. The size of the lesion was recorded by footpad swelling measurement every 10 days. RESULTS: The lesion growth was significantly hampered in treated mice compared with the untreated control group (P<0.05). Infrared radiation was more effective than microwave in inhibiting ulcer enlargement. CONCLUSION: Infrared radiation and microwave significantly hampered L. major lesion growth in BALB/c mice. This therapeutic effect was more in infrared radiation treated mice than microwave treated mice.

Is an occupation with vocal load a risk factor for laryngopharyngeal reflux: a prospective, multicentre, multivariate comparative study.

OBJECTIVES: To compare the prevalence of laryngopharyngeal reflux in choristers, teachers and control subjects without vocal load at work and to determine the risk factors for laryngopharyngeal reflux. DESIGN: A prospective, multicentre, multivariate comparative study. PARTICIPANTS: One hundred and nineteen singers from four professional choirs, seventy teachers from four schools and 111 control subjects with an occupation without vocal load. MAIN OUTCOME MEASURES: All participants completed a questionnaire about their dietary habits, height and weight, presence of stress, smoking and allergies, including the nine-item Reflux Symptom Index. The groups of choristers, teachers and the controls were compared with each other with regard to their sex, age, dietary habits, body-mass index, Reflux Symptom Index score and other factors affecting voice quality. The subjects from all three groups with suspected laryngopharyngeal reflux were compared with the subjects without it. The relationship between the Reflux Symptom Index score and the possible risk factors for reflux was estimated. RESULTS: The results showed significantly higher Reflux Symptom Index scores in the choristers than in the teachers and the controls (mean scores, 7.86 versus 6.33, P = 0.044; 7.86 versus 4.80, P = 0.000, respectively), but the number of subjects with suspected laryngopharyngeal reflux (Reflux Symptom Index score >13) did not differ significantly between the groups. The choristers were significantly more often treated for laryngopharyngeal reflux than the teachers and the controls (41%, 17% and 28%, respectively). The occupation chorister and frequently experiencing stress were the only factors that influenced the total Reflux Symptom Index score. CONCLUSIONS: Laryngopharyngeal reflux affects the choristers more often than the teachers or the control subjects without vocal load at work. These results suggest that singing as the main professional activity can notably contribute to the development of the reflux. Vocal load without singing is probably not an important aetiological factor for laryngopharyngeal reflux. The extraoesophageal symptoms affecting voice require treatment for laryngopharyngeal reflux and proper dietary habits especially in the group with high voice quality demands.

The role of nasogastric intubation on postoperative gastrointestinal function in patients with obstructive jaundice.

It is the practice of many surgeons to use the routine nasogastric tube after biliary operations, but its usefulness has been questioned. This study was designed to determine the effect of postoperative nasogastric intubation on gastrointestinal function in patients with obstructive jaundice. In this randomized clinical trial, 40 patients who underwent choledochoduodenostomy or hepaticojejunostomy were randomly divided into two groups. Patients in the experimental group did not have the nasogastric tube, and in the control group the nasogastric tube was routinely applied after surgery. Gastrointestinal function was compared in these two groups. Patients with no nasogastric intubation did not show any postoperative complications or prolonged hospital stay. On the contrary, nasogastric tube insertion postponed return of bowel function and increased the incidence of nausea and vomiting, while it did not affect the incidence of postoperative ileus. Routine use of the nasogastric tube after choledochoduodenostomy or hepaticojejunostomy can delay normal gastrointestinal function and increase postoperative discomfort.

High frequency of microsatellite instability in sporadic colorectal cancer patients in Iran.

Microsatellite instability in sporadic colorectal cancer patients was assessed, and the clinicopathological associations were evaluated in northeastern Iran, which is a high-risk region for gastrointestinal malignancies. Microsatellite instability (MSI) status of tumoral tissue, compared to normal tissue, was assessed with a standard panel of MSI markers on paraffin-embedded surgically resected tissues from 67 consecutive sporadic colorectal cancer patients. Eleven of the patients were under 40 years old. Female patients were significantly younger than male patients (mean age 54.2 vs 62.1 years, P = 0.020). MSI analysis revealed 18 cases of MSI-H (26.9%), 11 MSI-L (16.4%) and 38 MSS (microsatellite stable tumors; 56.7%). While a greater proportion of patients consisted of males, 56.7 vs 43.3% females, MSI-H was more frequent in females (34.5 vs 21.5%). MSI was associated with proximal location of tumor (P = 0.003) and lower stages of tumor (P = 0.002), while MSS tumors were associated with node metastasis. MSI has a higher frequency in sporadic colorectal cancer patients, suggesting that molecular epidemiology of the genetic alterations involved in colorectal cancer carcinogenesis has a different pattern in the Iranian population, which deserves further epidemiological attention. The high frequency of MSI-H in this population suggests that we should look at microsatellite instability prior to chemotherapy to determine the most appropriate chemotherapeutic strategy in our population.

Is C-reactive protein helpful for early diagnosis of acute appendicitis?

PURPOSE: Appendectomy is one of the most common surgical procedures all over the world. Although various laboratory tests and imaging studies are available to improve the accuracy of diagnosis, the rate of negative appendectomy is still about 15-30%. This study was designed to assess the diagnostic value of quantitative C-reactive protein (CRP) in patients suspected to acute appendicitis. MATERIALS AND METHODS: In a prospective study, blood samples of 102 patients were collected before appendectomy. CRP was measured by immunoturbidimetry and the data were compared with the final histopathologic reports. Diagnostic accuracy of the CRP test was analyzed by ROC curve. RESULTS: In histopathology, 83 patients (81/4%) had acute appendicitis and 19 (18/6%) had normal appendices. Considering 14 mg/lit as the cut-off point, this test shows 59% (95% CI, 48-69%) sensitivity and 68% (95% CI, 47-88%) specificity. The positive and negative predictive values were 89% (95% CI, 80-97%) and 27% (95% CI, 14-39%), respectively. CONCLUSIONS: The measurement of CRP levels is not an ideal diagnostic tool for ruling out or determination of acute appendicitis.

Uncut Roux-en-y esophagojejunostomy: A new reconstruction technique after total gastrectomy.

PURPOSE: Uncut Roux-en-y is a reconstruction method with a main purpose of diminishing Roux stasis syndrome. In this study we performed this procedure to evaluate utility and complications of this technique, as well as its effect on Roux stasis syndrome. METHODS: Total gastrectomy with Uncut Roux-en-y esophagojejunostomy was performed on 94 patients. This technique consists of an artificial jejunal occlusion 5 cm proximal to anastomosis and a jejunojejunostomy between afferent and efferent loops. Diagnosis of "Roux stasis syndrome" was made by clinical criteria. RESULTS: According to the mentioned criteria, the "Roux stasis syndrome" occurred in 21.2% of the patients. Whereas occurrence rate of other complications was: dysphasia (13.8%), benign stricture in anastomosis (9.6%), and fistula (5.3%). CONCLUSIONS: Comparing the results of our study to other related studies indicates that this type of operation has lower rate of "Roux stasis syndrome", and also decreases the postoperative stricture rate of the anastomosis.

SPINE workshop on automated X-ray analysis: a progress report.

The Structural Proteomics In Europe (SPINE) consortium contained a workpackage to address the automated X-ray analysis of macromolecules. The aim of this workpackage was to increase the throughput of three-dimensional structures while maintaining the high quality of conventional analyses. SPINE was able to bring together developers of software with users from the partner laboratories. Here, the results of a workshop organized by the consortium to evaluate software developed in the member laboratories against a set of bacterial targets are described. The major emphasis was on molecular-replacement suites, where automation was most advanced. Data processing and analysis, use of experimental phases and model construction were also addressed, albeit at a lower level.

Application of the use of high-throughput technologies to the determination of protein structures of bacterial and viral pathogens.

The Structural Proteomics In Europe (SPINE) programme is aimed at the development and implementation of high-throughput technologies for the efficient structure determination of proteins of biomedical importance, such as those of bacterial and viral pathogens linked to human health. Despite the challenging nature of some of these targets, 175 novel pathogen protein structures (approximately 220 including complexes) have been determined to date. Here the impact of several technologies on the structural determination of proteins from human pathogens is illustrated with selected examples, including the parallel expression of multiple constructs, the use of standardized refolding protocols and optimized crystallization screens.