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Enzyme-mediated processes have proven to be a valuable and sustainable alternative to traditional chemical methods. In this regard, the use of multi-enzymatic systems enables the realization of complex synthetic schemes, while also introducing a number of additional advantages, including the conversion of reversible reactions into irreversible processes, the partial or complete elimination of product inhibition problems, and the minimization of undesirable by-products. In addition, the immobilization of biocatalysts on magnetic supports allows for easy reusability and streamlines the downstream process. Herein we have developed a cascade system for cladribine synthesis based on the sequential action of two magnetic biocatalysts. For that purpose, purine 2′-deoxyribosyltransferase from Leishmania mexicana (LmPDT) and Escherichia coli hypoxanthine phosphoribosyltransferase (EcHPRT) were immobilized onto Ni2+-prechelated magnetic microspheres (MagReSyn®NTA). Among the resulting derivatives, MLmPDT3 (activity: 11,935 IU/gsupport, 63% retained activity, operational conditions: 40 °C and pH 5–7) and MEcHPRT3 (12,840 IU/gsupport, 45% retained activity, operational conditions: pH 5–8 and 40–60 °C) emerge as optimal catalysts for further synthetic application. Moreover, the MLmPDT3/MEcHPRT3 system was biochemically characterized and successfully applied to the one-pot synthesis of cladribine under various conditions. This methodology not only displayed a 1.67-fold improvement in cladribine synthesis (compared to MLmPDT3), but it also implied a practically complete transformation of the undesired by-product into a high-added-value product (90% conversion of Hyp into IMP). Finally, MLmPDT3/MEcHPRT3 was reused for 16 cycles, which displayed a 75% retained activity.

Fountoulakis K.N., Karakatsoulis G.N., Abraham S., Adorjan K., Ahmed H.U., Alarcón R.D., Arai K., Auwal S.S., Berk M., Bjedov S., Bobes J., Bobes-Bascaran T., Bourgin-Duchesnay J., Bredicean C.A., Bukelskis L., Burkadze A., Abud I.I.C., Castilla-Puentes R., Cetkovich M., Colon-Rivera H., Corral R., Cortez-Vergara C., Crepin P., De Berardis D., Delgado S.Z., De Lucena D., De Sousa A., Stefano R.D., Dodd S., Elek L.P., Elissa A., Erdelyi-Hamza B., Erzin G., Etchevers M.J., Falkai P., Farcas A., Fedotov I., Filatova V., Fountoulakis N.K., Frankova I., Franza F., Frias P., Galako T., Garay C.J., Garcia-Álvarez L., García-Portilla M.P., Gonda X., Gondek T.M., González D.M., Gould H., Grandinetti P., Grau A., Groudeva V., Hagin M., Harada T., Hasan T.M., Hashim N.A., Hilbig J., Hossain S., Iakimova R., Ibrahim M., Iftene F., Ignatenko Y., Irarrazaval M., Ismail Z., Ismayilova J., Jacobs A., Jakovljević M., Jakšić N., Javed A., Kafali H.Y., Karia S., Kazakova O., Khalifa D., Khaustova O., Koh S., Kopishinskaia S., Kosenko K., Koupidis S.A., Kovacs I., Kulig B., Lalljee A., Liewig J., Majid A., Malashonkova E., Malik K., Malik N.I., Mammadzada G., Mandalia B., Marazziti D., Marčinko D., Martinez S., Matiekus E., Mejia G., Memon R.S., Martínez X.E.M., Mickevičiūtė D., Milev R., Mohammed M., Molina-López A., Morozov P., Muhammad N.S., Mustač F., Naor M.S., Nassieb A., Navickas A., Okasha T., Pandova M., Panfil A.-L., Panteleeva L., Papava I., Patsali M.E., Pavlichenko A., Pejuskovic B., Da Costa M.P., Popkov M., Popovic D., Raduan N.J.N., Ramírez F.V., Rancans E., Razali S., Rebok F., Rewekant A., Flores E.N.R., Rivera-Encinas M.T., Saiz P., de Carmona M.S., Martínez D.S., Saw J.A., Saygili G., Schneidereit P., Shah B., Shirasaka T., Silagadze K., Sitanggang S., Skugarevsky O., Spikina A., Mahalingappa S.S., Stoyanova M., Szczegielniak A., Tamasan S.C., Tavormina G., Tavormina M.G.M., Theodorakis P.N., Tohen M., Tsapakis E.M., Tukhvatullina D., Ullah I., Vaidya R., Vega-Dienstmaier J.M., Vrublevska J., Vukovic O., Vysotska O., Widiasih N., Yashikhina A., Prezerakos P.E., Smirnova D.

Editorial of vol 32(1).Editorial del vol 32(1).

The main aim of this study was to determine the flexibility profile of the lower extremity in young taekwondo athletes, as well as identifying athletes with muscle shortness. For this purpose, a total of 17 young taekwondo athletes were assessed; seven from the Spanish team and ten from the Murcia team. The range of motion of the main movements of the lower extremity was evaluated by the "ROM SPORT" protocol and a descriptive analysis of each of the quantitative variables was performed. A Student t-test or arWilcoxon test was applied depending on the normality distribution of the data. The ROM values of each movement were classified as “normal” or “shortness” according to previously determined reference values. The results define the following flexibility profile: 23º for iliopsoas, 28º for piriformis, 37º for gastrocnemius, 42º for soleus, 46º for adductors, 46º for external rotators, 56º for internal rotators, 77º for monoarticular adductors, 80º for hamstrings, 140º for quadriceps and 145º for gluteus maximus. Individual evaluations identified muscle shortness in pyramidal (11 athletes, 64.7%), soleus (13 athletes, 76.4%), external rotators (8 athletes, 47%), monoarticular adductors (14 athletes, 82.3%), hamstrings (13 athletes, 76.4%), quadriceps (4 athletes, 23.5%) and gluteus maximus (12 athletes, 70.5%). This flexibility profile can help sports professionals to set quantifiable goals for the training of flexibility in this sport. The inclusion of stretching exercises or the increase of their doses should be applied for the improvement of the identified shortness in piriformis, soleus, external hip rotators, monoarticular, hamstring, quadriceps and gluteus maximus adductors.

[ES] El objetivo principal del presente estudio fue conocer el efecto de la categoría de edad sobre el perfil de flexibilidad en jóvenes taekwondistas usando pruebas angulares. Fueron reclutados un total de 17 taekwondistas jóvenes, que se clasificaron según su categoría federativa de edad en cadetes y junior. El rango de movimiento de los principales movimientos de la extremidad inferior fue valorado mediante el protocolo “ROM SPORT”. Se realizó un análisis descriptivo de cada una de las variables cuantitativas, que incluía la media y su correspondiente desviación típica. Una prueba t‐test fue empleada para determinar la existencia de diferencias entre la extremidad dominante y no dominante. Además, la misma prueba estadística analizó la interacción entre los 11 movimientos seleccionados y las categorías de edad. Por último, se calculó el tamaño del efecto de los resultados. Los resultados mostraron diferencias significativas entre ambas categorías en EC (psoas‐ilíaco), RIC (rotadores externos), REC (rotadores internos) y rotación total de la cadera (suma del RIC y REC). Estos resultados no coinciden con otros estudios que han valorado la flexibilidad en este mismo deporte. Los taekwondistas cadetes presentan valores de flexibilidad superiores a la categoría junior en los músculos rotadores internos y externo de cadera y en el psoas‐iliaco. [EN] The main aim of this study was to analyze the effect of the age category on the flexibility profile in young taekwondo athletes, by using angular tests. A total of 17 young taekwondo athletes were recruited, who were classified according to their federative age category. The range of motion of the main movements of the lower extremity was assessed by using the "ROM SPORT" protocol. A descriptive analysis was made of each of the quantitative variables, which included means and standard deviations. A t‐test was used to determine the existence of differences between the dominant and non‐dominant limbs. In addition, the same statistical test analyzed the interaction between the 11 selected movements and age categories. Finally, the effect size of the results was calculated. The results showed significant differences between cadets and juniors categories in EC (iliopsoas), RIC (external rotators), REC (internal rotators) and total rotation of the hip (sum of RIC and REC). These results are not in tune with other studies that have assessed flexibility in taekwondo. Cadet taekwondo athletes had higher values of flexibility than the junior category in the internal and external rotator muscles and in the psoas‐iliac.

The main aim of this study was to analyze the effect of the age category on the flexibility profile in young taekwondo athletes, by using angular tests. A total of 17 young taekwondo athletes were recruited, who were classified according to their federative age category. The range of motion of the main movements of the lower extremity was assessed by using the "ROM SPORT" protocol. A descriptive analysis was made of each of the quantitative variables, which included means and standard deviations. A t-test was used to determine the existence of differences between the dominant and non-dominant limbs. In addition, the same statistical test analyzed the interaction between the 11 selected movements and age categories. Finally, the effect size of the results was calculated. The results showed significant differences between cadets and juniors categories in EC (iliopsoas), RIC (external rotators), REC (internal rotators) and total rotation of the hip (sum of RIC and REC). These results are not in tune with other studies that have assessed flexibility in taekwondo. Cadet taekwondo athletes had higher values of flexibility than the junior category in the internal and external rotator muscles and in the psoas-iliac.

Studying Léon Werth's war diary, means uncovering the author's narrative identity in such extreme circumstances as the Occupation. Not only will this diary become a personal refuge in the face of a certain death, but also the space in which he will fight against the dissolution of his own ego. Thanks to the imbrication of events, thoughts and emotions, Werth is able to discover his Jewish identity. Furthermore, because of his commitment to writing, that identity will be fully formed, being aware of both the function and the mission of that same writing. Fighting silence, Werth understands that he is a surviving witness and that his testimony is a declaration under oath that appeals to the reader's sense of responsibility.

[EN] In 1940 occupied France, living conditions are so bad that the French find it difficult to survive. Nevertheless, however dreadful their material conditions, nothing can compare with the degradation of the soul’s life, the absence of the spiritual. The French suffer from hunger and cold, but, worse still, their soul dies. They lose their identity little by little. « The soul is today so void. We die of thirst », exclaimed Antoine de Saint-Exupéry in a letter to his mother dated 1940 (1994 : 83). Nevertheless, we cannot quench our thirst because there is only « this cesspool » (Saint-Exupéry, 1994 : 310). The pain is unbearable since it is the French spiritual patrimony that is at stake, a situation noted by numerous writers. The essence of life, the clear, clean water that grants life, turns into water of death, « a water that needs to go somber, a water that will absorb the dark suffering. All running water is destined to become slow, heavy. All running water is water about to die » (Bachelard, 1942 : 65-66). Life is choked, as explains Bachelard (1942 : 65), and there are only stagnant waters, « dead waters » (1942 : 63). In this paper I aim to show how writers feel and manifest this crisis, this loss of identity, this death ; and how they propose writing as a means of healing, as a way to salvation, particularly thanks to the autobiographical action. Writing will be the only place where they can quench their thirst, [ES] Cuando en 1940 Francia se convierte en territorio ocupado, las condiciones de vida son tan penosas que los franceses tienen grandes dificultades para sobrevivir. Sin embargo, por muy espantosas que sean las condiciones materiales, nada se puede comparar a la degradación de la vida del Espíritu, a la ausencia de lo espiritual. Los franceses pasan hambre, frío, pero peor aún, su espíritu muere. Los franceses van perdiendo su identidad. « El alma está tan yerma hoy. Morimos de sed », exclamaba Antoine de SaintExupéry en una carta a su madre en 1940 (1994 : 83)1 . Y sin embargo no podemos calmar nuestra sed porque solo existe « esta ciénaga » (Saint-Exupéry, 1994 : 310)2 El dolor es demasiado fuerte porque es el patrimonio espiritual francés el que está en peligro y numerosos escritores son conscientes de ello. La esencia de la vida, el agua clara, límpida, el agua que da la vida, se ha transformado en agua de muerte, « un agua que debe ensombrecerse, un agua que va a absorber el oscuro sufrimiento. Toda agua viva es un agua destinada a volverse lenta, pesada. Toda agua viva es un agua a punto de morir » (Bachelard, 1942 : 65-66). La vida queda ahogada como explica Bachelard (1942 : 65) y solo existen aguas estancadas, « aguas muertas » (1942 : 63)3 . En esta comunicación quiero mostrar cómo sienten y manifiestan los escritores esta crisis, esta pérdida de identidad, esta muerte ; y cómo proponen la escritura como remedio, como vía de salvación, gracias en especial a la acción autobiográfica. La escritura se convertirá en el único lugar en que poder calmar la sed., [FR] Quand en 1940 la France devient territoire occupé, les conditions de vie se dégradent à tel point que les Français vivent en grande difficulté. Pourtant, aussi épouvantables que les conditions matérielles de vie soient-elles, rien n’est comparable à la dégradation de la vie de l’Esprit, au manque du spirituel. Les Français sont affamés, les Français ont froid, mais pire encore, les Français meurent dans l’esprit. Les Français perdent peu à peu leur identité. « C’est l’âme aujourd’hui qui est tellement déserte. On meurt de soif » (1994 : 83), s’exclamait Antoine de Saint-Exupéry dans une lettre à sa mère en 1940. Et pourtant on ne peut pas s’abreuver parce qu’il n’existe que « ce marécage » (Saint-Exupéry, 1994 : 310). La douleur est trop forte parce que c’est le patrimoine spirituel français qui est en péril et de nombreux écrivains en sont très conscients. L’essence de la vie, l’eau claire, l’eau limpide, l’eau qui donne la vie, s’est transformée en eau de mort, « une eau qui doit s’assombrir, une eau qui va absorber la noire souffrance. Toute eau vive est une eau dont le destin est de s’alentir, de s’alourdir. Toute eau vivante est une eau qui est sur le point de mourir » (Bachelard, 1942 : 65-66). La vie est étouffée comme explique Bachelard (1942 : 65) et il n’y a que des eaux stagnantes, des « eaux mortes » (1942 : 63). Dans cette communication je voudrais montrer comment les écrivains ressentent et manifestent cette crise, cette perte d’identité, cette mort ; et comment l’écriture est proposée comme remède, comme voie de salut grâce à la démarche autobiographique d’une manière spéciale. L’écriture deviendra le seul endroit où aller pour s’abreuver.

Background Cognitive impairment is considered a central feature of psychotic disorders, with an important impact on prognosis and functional outcome (Nuechterlein et al., 2011). Among the etiological explanations on psychosis, several hypotheses involving alterations in the immune / inflammatory system have been proposed and recent research links these inflammatory processes with cognitive function, suggesting that the presence of inflammation is associated with poorer cognitive performance (Ribeiro-Santos et al., 2014, Cabrera et al., 2016). The study of the influence of gender on the possible association between inflammatory biomarkers and cognitive performance may favor the implementation of personalized treatments. The aim of the study is to investigate the association between inflammatory biomarkers and gender-based cognition in a sample of first psychotic episode (FEP). A total of 92 patients with a FEP, 62 men and 30 women, recruited in five clinical centers were included. A neurocognitive assessment was performed and the expression of pro and anti-inflammatory mediators in peripheral blood mononuclear cells (PBMC) and plasma was measured. Methods In the neurocognitive evaluation the domains of attention, verbal and working memory and executive function were included. Other and clinical variables included the assessment of psychotic and affective symptoms, age, sex, educational level and socio-economic level. The expression of the proinflammatory mediators (NFκB, iNOS, COX-2, PGE2, NO-2 and TBARS) and anti-inflammatory (15d-PGJ2, PPARγ and IκBα) of the main intracellular inflammatory pathway was measured in PBMC and plasma. Results A correlation was made between inflammatory biomarkers and neurocognitive performance according to gender, and significant associations were selected to perform a subsequent hierarchical regression analysis. In the final model, only the expression of COX2 was associated with better performance in executive function in males (B = 0.504, p =

En este articulo voy a mostrar como la funcion del pacto epistolar cohesiona la correspondencia intima familiar de Antoine de Saint-Exupery en el nivel pragmatico a traves de una clausula precisa como es la dimension dialogistica del intercambio epistolar. Como el pacto es un contrato en movimiento entre las instancias enunciativas necesita la presencia activa de las dos, no solo del epistolografo. El pacto es cumplido en la medida en que el destinatario responde a las cartas que el autor le envia. Cuanto mas constantes y regulares sean las respuestas, mas se asemejara el intercambio a un dialogo. El autor va creando esta dimension cuando hace de cada carta el espacio en el que los dialogos se dejan oir gracias a la inclusion de "concise analyses", "interior dialogue" y "fantasy dialogue", siguiendo a J. G. Altman. El dialogo epistolar deja de ser un lugar comun en las cartas de este autor.

This study, dealing with narrative identityduring the war in Henri Thomas’s epistolarydiscourse, implies the considerationof factors such as the awareness of his historicalself. Such awareness locates him inthe perspective of being a «story-man» thatcan be told in the discourse and becomeself-revealing, which situates us before aman liable to progress and for whom compromiseis his only aim. This will be thematerial used during the first stage of thewar (soldier Thomas), and it will later onbecome a spiritual compromise throughwhich he searches for his salvation and hiscountry’s, France. All this occurs thanks tothe creative silence through which life,identity, and literary universe appear inperfect communion.

Introduction The Mycobacterium tuberculosis complex includes the following species: Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, Mycobacterium bovis-BCG, Mycobacterium microti, Mycobacterium caprae, Mycobacterium pinnipedii, and Mycobacterium canettii. These species cause tuberculosis in humans and animals. Identification of mycobacterial strains has classically been performed by phenotype study. Over the last years, laboratories have developed several molecular techniques to differentiate between these species. The aim of this study is to evaluate these methods and develop a simple, fast, identification scheme. Material and methods We analyzed 251 strains randomly obtained from the strains studied in 2004, and 797 strains received by the Reference Laboratory between 2005 and 2007. Phenotype characterization of 4183 strains isolated during that period was done by studying the colony morphology, characteristics in culture, nitrate reduction, niacin accumulation, and growth in the presence of thiophen-2-carboxylic acid hydrazide 10 microg/mL and pyrazinamide 50 microg/mL. The molecular identification scheme designed was as follows: 1) gyrB PCR-RFLP with RsaI, TaqI or SacII and hsp65 RFLP/PCR with HhaI., and 2) multiplex-PCR to determine the presence/absence of the RD9 and RD1 regions. Results The results showed 100% agreement between phenotype study and the molecular scheme. Discussion This molecular identification scheme is a simple and fast method, with 100% sensitivity and specificity, that can be implemented in most clinical laboratories at a low cost.

The three-dimensional structure of a potent SSTR3-selective analogue of somatostatin, cyclo(3-14)H-Cys(3)-Phe(6)-Tyr(7)-D-Agl(8)(N(beta) Me, 2-naphthoyl)-Lys(9)-Thr(10)-Phe(11)-Cys(14)-OH (des-AA(1, 2, 4, 5, 12, 13)[Tyr(7), D-Agl(8)(N(beta) Me, 2-naphthoyl)]-SRIF) (peptide 1) has been determined by (1)H NMR in water and molecular dynamics (MD) simulations. The peptide exists in two conformational isomers differing mainly by the cis/trans isomerization of the side chain in residue 8. The structure of 1 is compared with the consensus structural motifs of other somatostatin analogues that bind predominantly to SSTR1, SSTR2/SSTR5 and SSTR4 receptors, and to the 3D structure of a non-selective SRIF analogue, cyclo(3-14)H-Cys(3)-Phe(6)-Tyr(7)-D-2Nal(8)-Lys(9)-Thr(10)-Phe(11)-Cys(14)-OH (des-AA(1, 2, 4, 5, 12, 13)[Tyr(7), D-2Nal(8)]-SRIF) (peptide 2). The structural determinant factors that could explain selectivity of peptide 1 for SSTR3 receptors are discussed.

Introduccion Mycobacterium kansasii se define como un patogeno oportunista que causa mayoritariamente enfermedad pulmonar. Esta especie representa el 9,7% de las micobacterias no tuberculosas identificadas en el Centro Nacional de Microbiologia durante los anos 2000-2003. Metodos En el presente estudio se analizan las caracteristicas fenotipicas y genotipicas de 298 cepasidentificadas como M. kansasii durante este periodo de tiempo. La caracterizacion fenotipica se realizo mediante metodos convencionales: caracteristicas morfologicas, de crecimiento y pruebas bioquimicas. La caracterizacion genotipica se realizo mediante amplificacion de un fragmento del gen hsp65 y posterior digestion con las enzimas BstEII y HaeIII segun el metodo descrito por Telenti. Resultados El 57,4% de las cepas estudiadas poseianlas caracteristicas fenotipicas propias de M. kansasii, pero en el resto se obtuvieron otros resultados que nos permitieron definir 17 biotipos. Se identificaron cepas pertenecientes a 6 de los 7 genotipos descritos, el 86,6% de las cepas se agrupaban en el genotipo I. Conclusion Al analizar las caracteristicas fenotipicas, se obtiene una mayor discriminacion para la diferenciacion intraespecifica de M. kansasii que al aplicar metodos genotipicos, pero debido al gran numero de cepas con caracteristicas fenotipicas atipicas, algunas de ellas muy especificas para la especie como la fotocromogenicidad, se aconseja la utilizacion de metodos convencionales y la aplicacion de metodos moleculares para la correcta identificacion de la especie.

In this study, we describe a multiplex PCR to detect a AGC→ACC (serine to threonine) mutation in the katG gene and a −15 C-to-T substitution ( inhA C−15T ) at the 5′ end of a presumed ribosome binding site in the promoter of the mabA - inhA operon. These mutations have been reported in the majority of previous studies as the most frequent mutations involved in the resistance to isoniazid (INH) of Mycobacterium tuberculosis clinical strains with high levels of resistance. The method was optimized and validated after an analysis of 30 M. tuberculosis clinical isolates with known sequences of the relevant part of the katG gene and the regulatory region of the mabA - inhA operon. We analyzed 297 INH-resistant M. tuberculosis isolates collected in Spain from 1996 to 2003 by PCR-restriction fragment length polymorphism (using the katG gene), DNA sequencing, and the newly developed multiplex PCR. The results were concordant for all 297 isolates tested. The analysis revealed that 204 (68.7%) of the isolates carried one or both of the mutations. This finding suggests that with further development this multiplex PCR will be able to detect the majority of the INH-resistant M. tuberculosis clinical isolates from Spain and other countries where a high frequency of similar mutations occur.

The Mycobacterium tuberculosis complex includes the following species: Mycobacterium tuberculosis, Mycobacterium africanum, Mycobacterium bovis, Mycobacterium bovis-BCG, Mycobacterium microti, Mycobacterium caprae, Mycobacterium pinnipedii, and Mycobacterium canettii. These species cause tuberculosis in humans and animals. Identification of mycobacterial strains has classically been performed by phenotype study. Over the last years, laboratories have developed several molecular techniques to differentiate between these species. The aim of this study is to evaluate these methods and develop a simple, fast, identification scheme.We analyzed 251 strains randomly obtained from the strains studied in 2004, and 797 strains received by the Reference Laboratory between 2005 and 2007. Phenotype characterization of 4183 strains isolated during that period was done by studying the colony morphology, characteristics in culture, nitrate reduction, niacin accumulation, and growth in the presence of thiophen-2-carboxylic acid hydrazide 10 microg/mL and pyrazinamide 50 microg/mL. The molecular identification scheme designed was as follows: 1) gyrB PCR-RFLP with RsaI, TaqI or SacII and hsp65 RFLP/PCR with HhaI., and 2) multiplex-PCR to determine the presence/absence of the RD9 and RD1 regions.The results showed 100% agreement between phenotype study and the molecular scheme.This molecular identification scheme is a simple and fast method, with 100% sensitivity and specificity, that can be implemented in most clinical laboratories at a low cost.

Mycobacterium kansasii is an opportunistic pathogen that mainly causes pulmonary infections. This species accounted for 9.7% of Mycobacteria other than tuberculosis complex identified in the reference laboratory of the Spanish Centro Nacional de Microbiologia during the period of 2000-2003.In this study we analyzed the phenotypic and genotypic characteristics of 298 M. kansasii strains isolated over this 4-year period. The phenotypic characteristics were determined by conventional methods: biochemical testing, culture and morphological study. Genotypic characteristics were studied using PCR restriction fragment analysis of a fragment of the hsp65 gene and digestion with BstEII and HaeIII, according to the method of Telenti.Among the total of tested strains, 57.4% had the typical phenotypic characteristics described for M. kansasii. The rest had atypical patterns that we grouped into 17 biotypes. Strains belonging to six of the seven described genotypes were identified, with 86.6% of the strains falling into genotype I.Analysis of the phenotypic characteristics of M. kansasii showed a higher discrimination index for intraspecific differentiation than genotypic methods. Nevertheless, the high variability of phenotypic characteristics, some of which were very specific for the species (e.g., photochromogenicity), could complicate their identification. Hence both conventional and molecular methods should be used to accurately identify the atypical isolates.