The different patterns of the caudal bursa of the Heligmonellidae (Nematoda) are redefined, taking into account the grouping of rays 2-6 and the sequence of origin of these rays from their common trunk. The type of symmetry of the caudal bursa is also redefined. The following patterns were observed and characterized: the basic patterns: types 2-3, 2-2-1, 1-3-1 and 1-4 and the intermediary patterns: type 2-3 tending to type 2-2-1, type 2-2-1 tending to type 1-3-1, type 1-3-1 tending to type 1-4 and type 2-2-1 tending to type 1-4. An evolutionary interpretation of the patterns is attempted and seems to follow the direction: 2-3 to 2-2-1 to 1-3-1 to 1-4. Seven atypical patterns are described. The caudal bursae were classified based on their symmetry: subsymmetrical, dissymmetrical and asymmetrical. Independently of the type of symmetry, the two latero-ventral lobes may have the same or different patterns. The type of symmetry, the ratio between the two latero-ventral lobes and a characteristic pattern were utilized to characterize the caudal bursae at the level of the genus and the subfamily. The combination of the right/left ratio and the type of symmetry gives heterogeneous results, with no real association between these characters. The most conspicuous asymmetries and dissymmetries were found among the Nippostrongylinae. The most frequent pattern in the Heligmonellidae is the basic type 2-2-1; types 1-3-1 and 1-4 are less frequent but are characteristic of several genera; type 1-4 is absent from the Heligmonellinae. Whatever the pattern, in the Heligmonellidae rays 4 and 5 are the last to diverge from the common trunk of rays 2-6. In 1981Heligmonina Baylis, 1928 and Stilestrongylus Freitas, Lent & Almeida, 1937, which do not show the same pattern in each lateral lobe. Within the framework of a revision of the Heligmonellidae, the aim of this work is to redefine the different patterns of the caudal bursa found within this family, taking into account not only the grouping of rays 2-6 but also the sequence of origin of these rays from their common trunk. The type of symmetry of the caudal bursa is also redefined. This enables us not only to make the descriptions of the caudal bursae more accurate, but also to highlight the characteristic type(s) of pattern within each genus and to attempt an evolutionary interpretation of the patterns.This descriptive system however seems to us to be insufficient to properly describe the caudal bursae of some Nippostrongylinae, especially the genera such as The elements usually considered in the description of a caudal bursa (CB) are: symmetry, pattern of latero-ventral lobes, development of the dorsal lobe and characters of the dorsal ray and rays 8. The characters of the dorsal ray and rays 8 are not treated in this work. Some terms concerning the symmetry and the pattern of the latero-ventral lobes are redefined as follows.Subsymmetrical bursae (CB SS): both lobes are of similar size and shape in relation to the axis of the dorsal ray, which passes through the sagittal axis of the worm .Fig. 2FDissymmetrical bursae (CB DS): One lobe is larger than the other in relation to the axis of the dorsal ray, which passes through the sagittal axis of the worm. When the right lobe (RL) is larger it is cited as CB DS RL+. When the left lobe (LL) is larger it is cited as CB DS LL+ .Asymmetrical bursae (CB AS): The axis of the dorsal ray does not pass through the sagittal axis but is displaced to the right or the left side of the worm. One lobe may or may not be larger than the other in relation to this axis. When the right lobe (RL) is larger it is cited as CB AS RL+. When the left lobe (LL) is larger it is cited as CB AS LL+ .There are three types, which concern the degree of development of the latero-ventral lobes in relation to the sagittal axis of the worm:Basic patterns: rays 2 to 6 show a characteristic arrangement of the following types: type 2-3 ; type 2-Intermediary patterns : rays 2 to 6 show an intermediary arrangement with features of two basic types: type 2-3 tending to type 2-2-1 (2-3 t 2-2-1) ; type 2-The main features considered in the definition of the patterns are the grouping of the rays and the sequence of origin (or divergence) of these rays from their common trunk. The level of divergence of a ray from a common trunk  is calculated on a straight line represented by ray 4; at the level of divergence another line is traced perpendicularly to the first one, as shown inThe data were compiled from descriptions published in the literature. We took into account only the caudal bursae where both latero-ventral lobes, or at least one, was spread out. In the latter case, the lobe was treated separately and the pattern was not extrapolated to the entire bursa.Durettestrongylus travassosi , Heligmonella vladimiri Sadovskaja, 1952, Heligmonina vogeli Khalil, 1931, Heligmonoides crassidorsualis Franco, 1967, Heligmonoides mirzai Smales, 2008, Heligmosomum deltaLongistriata degusi Babero & Cattan, 1975, Longistriata castrosilvai Almeida, 1934, Longistriata fortuita Freitas, Lent & Almeida, 1937, Longistriata perfidaGobindonema boodugi Sood & Parshad 1974, Morganiella cricetuli Yin & Zhang, 1981. They are here considered as Heligmonellidae incertae sedis since their generic allocation is not possible. In the particular case of H. crassidorsualis and H. mirzai, both were considered as Nippostrongylinae incertae sedis by Nesomystrongylus fissicauda Durette-Desset, Lehtonen & Haukisalmi, 2002. This species shares more characters with the Heligmosomidae than with the Heligmonellidae, i.e. presence of caudal spine in the female tail, caudal bursa with rays 2 and 3 well developed, small dorsal lobe, and axis of orientation of synlophe subfrontal. It is likely that the monospecific genus Nesomystrongylus Durette-Desset, Lehtonen & Haukisalmi, 2002 should be transferred to the Heligmosomidae.The systematic position of some species remains uncertain since, even if the caudal bursa is completely described, this is not the case with the synlophe. These species have therefore not been considered in this work. They are as follows: Rays 2 and 3 grouped, arising first and together from common trunk of rays 2 to 6.Rays 2 and 3 grouped from base in V-shape.Rays 4 to 6 having common trunk.Divergence of rays 4, 5 and 6 at same level.Type 2-3 Rays 2 ane.g. type 2-3 tending to type 2-2-1, see Material & Methods)  Rays 2 anRays 2 and 3 grouped, arising first and together from common trunk of rays 2 to 6.Ray 6 arising at the same level as ray 3.Distal divergence of rays 4 and 5.Type 2-2-1 Rays 2 anRays 2 and 6 arising first and at same level from common trunk of rays 2 to 6.Ray 3 arising just distally to level of divergence of rays 2 and 6.Distal divergence of rays 4 and 5.Type 2-2-1 t 1-3-1 Rays 2 anRays 2 and 6 arising first and at same level from common trunk of rays 2 to 6.Rays 3 to 5 having common trunk.Type 1-3-1 Rays 2 anRay 2 arising first from common trunk of rays 2 to 6.Ray 6 arising slightly distally to level of divergence of ray 2 and proximally to level of divergence of ray 3.Rays 3 to 6 having short common trunk.Type 1-3-1 t 1-4 Ray 2 ariRay 2 arising first from common trunk of rays 2 to 6.Rays 3 to 6 having long common trunk.Type 1-4  (Brevistriatinae) and MalvinemaMalvinema, M. carolinaeM. victoriaeM. scapteromys  and M. yagoi Digiani & Durette-Desset, 2003, show the type 1-4; in two species of Srivastavanema, S. cynocephali Durette-Desset & Purwaningsih, 1999 and S. yapi Durette-Desset & Lim Boo Liat, 1975, both lobes are of type 2-3 whereas a third species S. bhagwansinghi Durette-Desset & Lim Boo Liat, 1975 shows a type 1-4 in both lobes. We thus consider this type 2-3 as different from that of the most basal species. It may be derived from the type 1-4 by the migration of ray 3 towards ray 2, whereas rays 4 to 6 still have a long common trunk.A particular 2-3 type was observed in some bursae or isolated lobes of the genera Cordicauda cordicauda  and N. marhaeniaeType 1-3-1 t 4-1 , RL is pNippostrongylus sembeli Hasegawa & Tarore, 1995; and in the left lobes of N. magnus, N. rauschi Chabaud & Desset, 1966 and N. typicus . This type seems directly derived from type 1-3-1, involving a marked distal migration of ray 2 towards ray 3. Ray 6 is the most proximal ray to diverge from the common trunk of rays 2-6, and rays 2 to 5 form a long common trunk on which ray 2 arises distinctly distally.Type 4-1 , LL is pNippostrongylus: N. djumachani , N. marhaeniae, N. rysavyi  and N. witenbergi Greenberg, 1972. It seems directly derived from type 4-1 by the migration of ray 5 from its base towards ray 6. Ray 5 diverges from the common trunk at the same level as ray 6 and there is persistence of the common trunk of rays 2 to 4.Type 3-1-1 , LL is pSciuricola dremomys  and S. moreli  (Heligmonellinae). It seems directly derived from type 1-3-1 by the migration (but not apposition) of ray 5 towards ray 6, with persistence of the group formed by rays 3 and 4.Type 1-2-1-1 , LL is pTricholinstowia maseriTricholinstowia) by the migration of ray 5 towards ray 6. Type 2-1-2 ; dissymmetrical with right lobe larger (CB DS RL+); dissymmetrical with left lobe larger (CB DS LL+); asymmetrical (CB AS); asymmetrical with right lobe larger (CB AS RL+); asymmetrical with left lobe larger (CB AS LL+). At the generic level, we consider that a given genus may be characterized by the type of symmetry most frequently found among the species belonging to this genus.As mentioned above, a species can have the same or a different pattern in both latero-ventral lobes. On the other hand, as seen above, the different patterns may be interpreted from an evolutionary point of view. This determines three types of “evolutionary ratio” between both lateral lobes (“right/left ratio”) in a caudal bursa: (1) the same pattern in both lobes; (2) the pattern of the right lobe is derived with respect to that of the left lobe (“RL derived”); or (3) the pattern of the left lobe is derived with respect to that of the right lobe (“LL derived”).Sciurodendrium Durette-Desset, 1971  we propose that the characteristic pattern is of type 2-2-1 because amongst the 11 treatable lobes, four are of type 2-2-1, three of type 2-3 t 2-2-1, and two of type 2-2-1 t 1-3-1. Some genera may have two characteristic patterns. For example in Heligmonina  two characteristic patterns are proposed: 1-3-1 and 1-4; type 1-3-1 being the most frequent in the right lobe and type 1-4 in the left lobe (21 and 19 lobes respectively).Within a given genus the bursal pattern usually varies among the species. Moreover, as seen immediately above, most species have a different pattern in each lobe. This means that in some genera we can find several different patterns. However it seems possible to choose one or two characteristic patterns for each genus. The characteristic pattern is determined by the type most frequently found, which is here interpreted as the most frequent basic type plus the contiguous intermediary types. For example, for the genus Heligmonellidae  Durette-Desset, 1971 Heligmonellinae  Durette-Desset & Chabaud, 1977 Heligmonella Mönnig, 1927 (3/4)CB SS (2 spp.) or CB DS RL+ (1 sp.). CB with the same pattern in both lobes (2 spp.) or with LL derived (1 sp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 1-4. Characteristic pattern: 2-2-1.Paraheligmonella Durette-Desset, 1971 (4/5)CB SS. CB with same pattern in both lobes (3 spp.) or with RL derived (1 sp.). Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1, atypical 2-2-1 t 4-1. Characteristic pattern: 2-2-1.Paraheligmonella cubaensis , with the pattern 1-4 in both lobes was considered with reservations as belonging to Paraheligmonella CB SS. CB with different patterns in both lobes (1 sp.). Patterns observed: 2-3 t 2-2-1, atypical 1-2-1-1. Characteristic pattern: the data are insufficient to choose a characteristic pattern but two characters of the CB shared by both species are rays 4 and 5 separated from base and dorsal lobe distinct.Tricholinstowia Travassos, 1937 (8/8)CB SS (6 spp.) or CB DS LL+ (1 sp.), type of symmetry unknown in 1 sp. CB with the same pattern in both lobes (4 spp.) or with RL derived (1 sp.) or with LL derived (1 sp.), right/left ratio unknown in 2 spp. Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 1-3-1, atypical 1-2-2 and 2-1-2. Characteristic pattern: 1-3-1; rays 5 and 6 pincer-shaped.T. mogera  and T. morenishi .Remark: only the right lobe is treatable in Trichotravassosia Lent & Freitas, 1938 (2/2)CB SS. CB with the same pattern in both lobes. Patterns observed: 2-2-1 t 1-4, atypical 2-2-1 t 4-1. Characteristic pattern: 2-2-1.T. capromydis Baruš & Rysavý, 1967 only the right lobe is treatable, showing an atypical pattern of type 2-2-1 tending to 4-1.Remark: in XericolaCB SS. CB with the same pattern in both lobes. Patterns observed: 2-2-1 t 1-4. Characteristic pattern: 2-2-1.Brevistriatinae Durette-Desset, 1971 Brevistriata Travassos, 1937 (3/4)CB SS. CB with the same pattern in both lobes. Patterns observed: 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.B. fukiensis Wang, Zao & Chen, 1978, is not included as the published illustration is not of a fully opened bursa.Remark: the caudal bursa of Calypsostrongylus Schmidt, Myers & Kuntz, 1967 (7/7)CB SS (5 spp.) or CB DS RL+ (2 spp.). CB with the same pattern in both lobes (5 spp.) or with LL derived (2 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-4. Characteristic pattern: 2-2-1.Cordicauda Durette-Desset, 1971 (5/5)CB DS LL+ (3 spp.) or CB SS (2 spp.). CB with the same pattern in both lobes (3 spp.) or LL derived (2 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 1-4. Characteristic pattern: 2-2-1.Fissicauda Durette-Desset & Krishnansamy, 1976 (6/6)CB SS (3 spp.) or CB DS LL+ (2 spp.) or CB DS RL+ (1 sp.). CB with the same pattern in both lobes (3 spp.) or with RL derived (3 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 1-3-1. Characteristic pattern: 2-2-1.Kuala Durette-Desset & Krishnansamy, 1976 (2/2)CB SS. CB with the same pattern in both lobes. Pattern observed: atypical type 1-2-2.Lagostrongylus Fukumoto, Kamiya & Ohbayashi, 1986 (3/3)CB SS. CB with the same pattern in both lobes. Patterns observed: 2-3 t 2-2-1. Characteristic pattern: 2-3 t 2-2-1.Macrostrongylus Ow-Yang, Durette-Desset & Ohbayashi, 1983 (2/2)CB SS. CB with the same pattern in both lobes. Patterns observed: 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.Metheligmonella Durette-Desset, 1971 (2/2)CB SS. CB with the same pattern in both lobes. Patterns observed: 2-3 t 2-2-1. Characteristic pattern: 2-3 t 2-2-1.Paraheligmonina  (27/28)CB SS (12 spp.) or with a slight left (8 spp.) or right (7 spp.) dissymmetry. CB with the same pattern in both lobes (18 spp.) or with LL derived (7 spp.) or with RL derived (2 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 1-3-1, 1-4, atypical 2-2-1 t 4-1 and 4-1. Characteristic patterns: 2-2-1, 1-3-1.P. trifurcata  is not included as the published illustration is not of a fully opened bursa.Remark: the caudal bursa of QuentinstrongylusCB SS. CB with the same pattern in both lobes. Patterns observed: 1-3-1. Characteristic pattern: 1-3-1.Srivastavanema  (3/5)CB SS. CB with the same pattern in both lobes. Patterns observed: secondary 2-3, and 1-4. Characteristic pattern: 1-4.Pudicinae  Durette-Desset, 1971 Acanthostrongylus Travassos, 1937 (1/1)CB SS. CB with the same pattern in both lobes. Pattern observed: 1-4. Characteristic pattern: 1-4.Durettestrongylus Guerrero, 1982 (2/3)CB SS. CB with the same pattern in both lobes. Patterns observed: 2-2-1. Characteristic pattern: 2-2-1.Freitastrongylus Gonçalves, Pinto & Durette-Desset (1/1)CB SS. CB with the same pattern in both lobes. Pattern observed: 1-4. Characteristic pattern: 1-4.Fuellebornema Travassos & Darriba, 1929 (6/7)CB SS. CB with the same pattern in both lobes. Pattern observed: 1-4. Characteristic pattern: 1-4.F. almeidai Travassos, 1937 is not included as the published illustration is not of a fully opened bursa.Remark: the caudal bursa of Heligmostrongylus Travassos, 1917 (9/9)CB SS. CB with the same pattern in both lobes (8 spp.) or with LL derived (1 sp.). Patterns observed: 2-2-1, 2-2-1 t 1-3-1, 1-3-1. Characteristic pattern: 2-2-1.Justinema R’Kha & Durette-Desset, 1991 (3/3)CB SS. CB with RL derived (2 spp.) or with the same pattern in both lobes (1 sp.). Patterns observed: 2-2-1, 2-2-1 t 1-3-1. Characteristic pattern: 2-2-1.Pseudoheligmosomum Travasssos, 1937 (1/1)CB AS LL+. CB with the same pattern in both lobes. Pattern observed: 2-3. Characteristic pattern: 2-3.Pudica Travassos & Darriba, 1929 (12/12)CB SS. CB with the same pattern in both lobes (11 spp.) or with RL derived (1 sp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 1-3-1, 1-4. Characteristic patterns: 2-2-1, 1-3-1.Sciurodendrium Durette-Desset, 1971 (6/7)CB SS. CB with LL derived (3 spp.) or with the same pattern in both lobes (2 spp.), right/left ratio unknown in 1 sp. Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 1-3-1. Characteristic pattern: 2-2-1.S. aripense  is not included as the published illustration is not of a fully opened bursa. Only the right lobe is treatable in S. bravohollisae Falcón-Ordaz & Lamothe-Argumedo, 2006.Remarks: the caudal bursa of Nippostrongylinae Durette-Desset, 1971 Bunomystrongylus Hasegawa & Mangali, 1996 (2/2)CB DS RL+ (1 sp.) and CB DS LL+ (1 sp.). CB with RL derived. Patterns observed: 2-3 t 2-2-1, 2-2-1, 1-3-1, 1-4. No characteristic pattern.Carolinensis  (12/12)CB SS (7 spp.) or CB DS LL+ (4 spp.) or CB DS LL+ (1 sp.). CB with the same pattern in both lobes (9 spp.) or with RL derived (1 sp.) or with LL derived (1 sp.), right/left ratio unknown in one species. Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 1-3-1, 1-3-1 t 1-4, 1-4. Characteristic pattern: 2-2-1.C. eothenomysi Asakawa, Kamiya & Ohbayashi, 1986 and C. huehuetlana Falcón-Ordaz & Sanabria Espinosa, 1996, the left lobe being not spread out.Remark: only the right lobe is treatable in Euzetoda Elias & Durette-Desset, 2003 (1/1)CB SS. CB with the same pattern in both lobes. Pattern observed: 2-3 t 2-2-1. Characteristic pattern: 2-3 t 2-2-1.Guerrerostrongylus Sutton & Durette-Desset, 1991 (2/2)CB SS. CB with the same pattern in both lobes. Patterns observed: 1-3-1. Characteristic pattern: 1-3-1.Hasanuddinia Hasegawa & Syafruddin, 1994 (1/1)CB SS. CB with RL derived. Patterns observed: 2-2-1 t 1-3-1, 2-2-1 t 1-4. Characteristic pattern: 2-2-1.Hassalstrongylus Durette-Desset, 1971 (13/14)H. mazzai Freitas, Lent & Almeida, 1937 is not included as the published illustration is not of a fully opened bursa.CB SS (6 spp.) or DS RL+ (6 spp.) or DS LL+ (1 sp.). CB with the same pattern in both lobes (9 spp.) or RL derived (2 spp.) or LL derived (2 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 1-3-1, 1-4. Characteristic patterns: 1-4 (type species), 2-2-1, 1-3-1. Remark: the caudal bursa of Heligmonina Baylis, 1928 (24/26)CB DS LL+. CB with LL derived (21 spp.) or with the same pattern in both lobes (3 spp.). Patterns observed: 1-3-1, 1-3-1 t 1-4, 1-4. Characteristic patterns: 1-3-1 on right lobe, 1-4 on left lobe.H. oenomyos Baylis, 1928 is not illustrated and therefore untreatable. The left lobe of H. cricetomyos Baylis, 1928 is typical of the genus Heligmonina i.e. of type 1-4, but the right lobe is atypical with a type 2-2-1 not retained in the patterns observed.Remarks: the caudal bursa of Heligmonoides Baylis, 1928 (12/12)CB DS LL+. CB with the same pattern in both lobes (6 spp.) or with RL derived (3 spp.) or LL derived (3 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1- 3-1, 1-3-1, 1-4. Characteristic pattern: 2-2-1.Hypocristata Durette-Desset, 1971 (3/3)CB SS (2 spp.) or DS RL+ (1 sp.). CB with the same pattern in both lobes (1 sp.) or with RL derived (1 sp.) or with LL derived (1 sp.). Patterns observed: 2-2-1, 2-2-1 t 1-4, 2-2-1 t 1-3-1, 1-3-1. Characteristic pattern: 1-3-1.Malaistrongylus Ow-Yang, Durette-Desset & Ohbayashi, 1983 (1/1)CB SS. CB with the same pattern in both lobes. Pattern observed: 2-2-1. Characteristic pattern: 2-2-1.MalvinemaCB DS RL+ (3 spp.) or CB AS RL+ (1 sp.). CB with the same pattern in both lobes (2 spp.) or with RL derived (1 sp.) or with LL derived (1 sp.). Patterns observed: 1-4, secondary 2-3. Characteristic pattern: 1-4.Mammanidula Sadovskaja, 1952 (3/5)CB AS RL+ (2 spp.) or CB SS (1 sp.). CB with LL derived (2 spp.) or RL derived (1 sp.). Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.M. melomyos  and M. siamensis Ohbayashi & Vajrasthiva, 1983 are not included as the published illustrations are not of fully opened bursae.Remark: the caudal bursae of MawsonemaCB SS. CB with the same pattern in both lobes. Pattern observed: 2-3. Characteristic pattern: 2-3.Maxomystrongylus Hasegawa & Syafruddin, 1997 (2/2)CB DS LL+. CB with the same pattern in both lobes (1 sp.) or with RL derived (1 sp.). Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.Melomystrongylus Smales, 2009 (2/2)CB DS LL+ or CB DS RL+. CB with RL derived (1 sp.), right/left ratio unknown in one species. Patterns observed: 2-3, 1-3-1. Characteristic pattern: 2-3.M. sepikensis Smales, 2009 is illustrated and treatable.Remark: only the left lobe of MontistrongylusCB DS RL+. CB with RL derived. Patterns observed: 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.Neoheligmonella Durette-Desset, 1971 (21/23)N. affinis  is not treatable since it is not illustrated in the original description. Only the right lobe is treatable in N. impudica  and N. moennigi . N. lemniscomysi  is the only species having a caudal bursa with a right lobe larger and with the same pattern (1-3-1) in both lobes. On the other hand, its synlophe is also very different from that of the remaining species in the genus. It is likely that this species belongs to a different genus and it is not treated herein.CB SS. CB with the same pattern in both lobes (for at least 12 spp.) or with RL derived (for at least 7 spp.), right/left ratio unknown in 2 spp. Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-3 t 2-2-1. Remarks: the caudal bursa of Nippostrongylus Lane, 1923 (9/9)CB AS RL+ (6 spp.) or CB DS RL+ (3 spp.). CB with LL derived. Patterns observed: 1-3-1, 1-3-1 t 1-4, 1-3- 1 t 4-1, 4-1, 3-1-1. Characteristic patterns: 1-3-1 and derived atypical 4-1 on right lobe; atypical 4-1 and derived atypical 3-1-1 on left lobe.Odilia Durette-Desset, 1973 (18/19)CB SS (8 spp.) or DS RL+ (3 spp.) or DS LL+ (2 spp.), symmetry unknown in 5 spp. CB with same pattern in both lobes (at least 8 spp.) or with RL derived (at least 3 spp.), right/left ratio unknown in 5 spp. Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1, 2-2-1 t 1-3-1, 2-2-1 t 1-4, 1-3-1, 1-3-1 t 1-4, 1-4. Characteristic patterns: 2-2-1, 1-4.O. polyrhabdote  is not included as the published illustration is not of a fully opened bursa. Only one lobe is treatable in O. brachybursa , O. implexa Smales, 2008, O. moatensis , O. similis Smales, 2009 and O. uromyos .Remarks: the caudal bursa of Orientostrongylus Durette-Desset, 1970 (7/8)CB SS. CB with the same pattern in both lobes (6 spp.) or with LL derived (1 sp.). Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.O. siamensis Ohbayashi & Kamiya, 1980 is not included as the published illustration is not of a fully opened bursa.Remark: the caudal bursa of Paraheligmonelloides Fukumoto, Kamiya & Suzuki, 1980 (9/9)CB DS LL+ (4 spp.) or CB DS RL+ (3 spp) or CB SS (2 spp.). CB with the same pattern in both lobes. Patterns observed: 2-3, 2-3 t 2-2-1, 2-2-1, 1-4. Characteristic patterns: 2-2-1, 1-4.P. singauwaensis Smales, 2009 is treated from its redescription CB SS (1 sp.) or CB DS LL+ (1 sp.). CB with the same pattern in both lobes. Patterns observed: 2-2-1, 2-2-1 t 1-3-1. Characteristic pattern: 2-2-1.Sabanema Ow-Yang, Durette-Desset & Ohbayashi, 1983 (4/5)CB DS RL+ (3 spp.) or CB SS (1 sp.). CB with the same pattern in both lobes (2 spp.) or with RL derived (1 sp.) or with LL derived (1 sp.). Patterns observed: 2-3 t 2-2-1, 2-2-1. Characteristic pattern: 2-2-1.S. kepongi Ow Yang, Durette-Desset & Ohbayashi, 1983 is not included as the published illustration of the bursa is not of a fully opened bursa.Remark: the caudal bursa of SpalacinaCB SS (2 spp.) or DS LL+ (1 sp.). CB with the same pattern in both lobes (at least 2 spp.), right/left ratio unknown in one species. Patterns observed: 1-3-1 t 1-4, 1-4. Characteristic pattern: 1-4.S. spalacis  the left lobe being not spread out.Remark: only the right lobe is treatable in Stilestrongylus Freitas, Lent & Almeida, 1937 (24/25)CB DS RL+ (21 spp.) or CB SS (3 spp.). CB with RL derived (11 spp.) or with the same pattern in both lobes (10 spp.) or with LL derived (3 spp.). Patterns observed: 2-3 t 2-2-1, 2-2-1, 1-3-1, 1-4. Characteristic patterns: 2-2-1, 1-4.S. peromysci Falcón-Ordaz & Sanabria-Espinoza, 1999 is not included as the published illustration is not of a fully opened bursa.Remark: the caudal bursa of Suttonema Digiani & Durette-Desset, 2003 (1/1)CB DS RL+. CB with the same pattern in both lobes. Pattern observed: 1-4. Characteristic pattern: 1-4.Trichofreitasia Sutton & Durette-Desset, 1991 (1/1)CB SS. CB with the same pattern in both lobes. Pattern observed: 2-2-1. Characteristic pattern: 2-2-1.Yatinema Asakawa & Ohbayashi, 1985 (2/2)CB SS (1 sp.) or CB DS RL+ (1 sp.). CB with RL derived (1 sp.), right/left ratio unknown in one species. Patterns observed: 2-2-1 t 1-4, 1-3-1. Characteristic pattern: 2-2-1.Y. siamensis Asakawa, Kamiya & Ohbayashi, 1986 is treatable, the left lobe being not spread out.Remark: only the right lobe of Based on the elements considered above, the four subfamilies may be synthetically characterized as follows: the Heligmonellinae by a subsymmetrical caudal bursa, with the same pattern in both lobes, and a characteristic pattern of type 2-2-1 ; the BreOf the five main patterns recognized among the Trichostrongylina treated by In the present article, we provide an additional feature at the descriptive level, which is the sequence of the origin of the rays from the common trunk. Consequently, the presence of a new basic pattern (type 1-4) is highlighted, plus the presence of intermediary types: type 2-3 tending to 2-2-1, type 2-2-1 tending to 1-3-1, type 1-3-1 tending to 1-4 and type 2-2-1 tending to 1- 4, which have enabled us to attempt an evolutionary interpretation of the patterns.Type 2-3, the basal type, is somewhat infrequent, but not the following intermediary type, 2-3 t 2-2-1, which is common and present in all four subfamilies. The basic type, 2-2-1, seems to be the most consistent pattern in the family: it is the most frequent characteristic pattern, is present in all four subfamilies, and there are relatively few intermediary types from 2-2-1 to 1-3-1, to 1-4 or to atypical patterns. Types 1-3-1 and 1-4 are less frequent than type 2-2-1 but are characteristic of several genera; type 1-3-1 is observed in all four subfamilies, and is the origin of several atypical types ; type 1-4, characteristic of laterally elongated lobes, is absent from the Heligmonellinae. Finally, the family Heligmonellidae is characterized by the fact that, regardless of the pattern, with very few exceptions, rays 4 and 5 are always the last to diverge from the common trunk of rays 2 to 6.It is clear that a dissymmetrical or asymmetrical caudal bursa should be regarded as derived with respect to a subsymmetrical one. However, the type of symmetry has been actually considered a character of little value above the species level, since the asymmetrical bursae and mostly the dissymmetrical bursae have arisen several times during the course of evolution. The dissymmetry usually involves the transverse  lengthening of one of the lobes. This dissymmetry probably plays a role in holding the female during copulation . The mose.g. Neoheligmonella 2-3 t 2-2-1 and 2-2-1; Heligmonina 1-3-1 and 1-4); there are very few cases in which both lobes have a totally different pattern .Independent of the type of symmetry, both lobes of the caudal bursa may have the same or different patterns. In the latter case the most frequent situation from an evolutionary point of view is one lobe having one pattern and the other lobe showing the “next” intermediary or basic type , or it is always the same lobe, which is derived with respect to the other (Heligmonina). Other genera are less homogeneous and the right/left ratio varies among the species of the genus  or the basal pattern . In genera with subsymmetrical or slightly dissymmetrical caudal bursae, not only may the right/left ratio vary but also the type of symmetry within each genus .The combination of the right/left ratio and type of symmetry gives disparate results: in genera with marked dissymmetry or asymmetry, the hypertrophied lobe is usually the same but it may show either the derived pattern . The Nippostrongylinae, in which the symmetry and the pattern in both lobes are more variable, are parasitic in a group, which appeared more recently, the muroid rodents (mainly cricetids and murids).


Effective in vivo protection from Shiga toxin Type 1 (Stx1) is achieved by polymer-bound, heterobifunctional inhibitors-adaptors (PolyBAITs), which exhibit prolonged half-life in circulation and by mediating formation of face-to-face SAP-AB5 complexes, block receptor recognition sites and redirect toxins to the spleen and liver for degradation. Direct correlation between solid-phase activity and protective dose of PolyBAITs both in the cytotoxicity assay and in vivo indicate that the mechanism of protection from intoxication is inhibition of toxin binding to the host cell membrane. The polymeric scaffold influences the activity not only by clustering active binding fragments but also by sterically interfering with the supramolecular complex assembly. Thus, inhibitors based on N-(2-hydroxypropyl) methacrylamide (HPMA) show significantly lower activity than polyacrylamide-based analogs. The detrimental steric effect can partially be alleviated by extending the length of the spacer, which separates pendant ligand from the backbone, as well as extending the spacer, which spans the distance between binding moieties within each heterobifunctional ligand. Herein we report that polymer size and payload of the active ligand had moderate effects on the inhibitor’s activity.Inhibition of AB Escherichia coli (STEC), particularly the O157:H7 strain, is the leading cause of hemolytic-uremic syndrome (HUS) in industrialized countries  of the glycolipid, Gb3. k, designed to sequester Stx in the intestine before Stx entered the circulation. Unfortunately, this approach failed to demonstrate significant efficacy in a Phase 2 clinical trial ethylcarbamate +.arbamate  carbamoyl-4-O--1,2-O-[(S)-1-(methoxycarbonyl)ethylidene]-α-D-glucopyranoside (3).3- Alkyne 1  was dissolved in TFA (1.6 mL) and left at room temperature for 1 h. The mixture was concentrated, treated with Et3N and concentrated again then 3-O-acetyl-6-O-(4-nitrophenyloxycarbonyl)-4-O--1,2-O-[(S)-1-(methoxycarbonyl) ethylidene]-α-D-glucopyranoside 2 +.O-[(S)-1-(carboxy)ethylidene]-4-O--6-O-carbamoyl-α-D-glucopyranoside (4).1,2- Disaccharide 3  was dissolved in dry methanol (4 mL) and 1 M NaOMe (0.54 mL) was added. The solution was stirred at room temperature for 16 h. The mixture was concentrated and dissolved in water. After 0.5 h TLC indicated that hydrolysis of the methyl ester was complete. The mixture was neutralized with 5 M acidic acid and lyophilized to provide the product 4 as white foam . 1H-NMR (D2O) δ: 5.62 , 4.67 , 4.44 , 4.41–4.37 , 4.24 , 4.18 , 4.06–4.01 , 3.91 , 3.82–3.60 , 3.55 , 3.36–3.32 , 2.9 , 1.64 . ESI HRMS: m/z: calcd for C26H39N2O18: 667.2203; found 667.2205 [M − H]−.O-[(S)-1-(carboxy)ethylidene]-4-O1,2-O--βO-carbamoyl-α-D-galactopyranosyl]-6--D-glucopyranoside (5). Compound 4  was dissolved in water (5.34 mL) and the pH was adjusted to 7.5 by addition of solid NaHCO3. Then 1 M HEPES buffer  was added followed by 0.4 M DTT solution (500 μL) and alkaline phosphatase . UDP-glucose  was dissolved in the reaction mixture and the fusion enzyme GalT/UDP-4'-epimerase −.pimerase  (950 μL)R,S)-5-(3-azido-2-hydroxypropoxy)-N,N,2-trimethyl-1,3-dioxane-2-carboxamide (7). to provide the title compound 7 as a clear syrup . 1H-NMR (CDCl3) δ: 4.10–4.06 , 3.92–3.82 , 3.70–3.48 , 3.40–3.28 , 3.24 and 3.02 , 1.52 ; ESI HRMS: m/z: calcd for C11H20N4O5Na: 311.1326; found: 311.1329 [M + Na]+.poxide 6  -5-{3-azido-2-propoxy}-N,N,2-trimethyl-1,3-dioxane-2-carboxamide (9). and lactosyl trichloroacetimidate 8 +.midate 8  -5-{3-acetamido-2-propoxy}-N,N,2-trimethyl-1,3-dioxane-2-carboxamide (10). in dry THF (19 mL) triphenylphosphine  was added and the mixture was stirred at 65 °C for 6 h then water (2.5 mL) was added and the mixture was stirred at 65 °C for 6.5 h, then concentrated, co-evaporated with toluene and dried under vacuum. The residue was dissolved in pyridine (20 mL) and acetic anhydride (10 mL) was added then the mixture was left at room temperature overnight. The mixture was concentrated, co-evaporated with toluene and the residue was chromatographed on silica gel using hexane-acetone (1:1) to provide N-acetate 10 as a white foam ; 1H-NMR (CDCl3) δ: 6.07 , 5.89 , 5.35 , 5.18 , 5.13–5.10 , 4.98 , 4.88–4.83 , 4.63 , 4.60–4.56 , 4.55 , 4.50 , 4.16–4.02 , 3.90–3.86 , 3.79–3.70 , 3.66–3.41 , 3.22 , 3.21–3.12 , 2.17–2.04 , 1.96 , 1.50 ; ESI HRMS: m/z: calcd for C39H58N2O23Na: 945.3323, found: 945.3315 [M + Na]+. R,S)-5-{3-(prop-2-yn)oxycarbonylamino-2-[4--β-D-glucopyranosyloxy]propoxy}-2-methyl-1,3-dioxane-2-carboxylic acid (12). was dissolved in methanol (4 mL) and transferred to a plastic Falcon tube, then 4 M NaOH  was added. The mixture was stirred at 80 °C. After 3 days the reaction was not complete as checked by NMR. More 4 M NaOH  was added and stirring was continued overnight at 85 °C. The mixture containing hydrolysis product 11 was treated with dry ice until the solution reached pH 8–9. Then propargyl chloroformate  was added and the mixture was stirred at room temperature. After 2 h the mixture was purified on HPLC (C-18) using water containing 0.1% TFA—acetonitrile gradient. Two fractions were isolated, concentrated and freeze-dried. First fraction was the product of incomplete hydrolysis of 10 -5-{3-acetamido-2-[4--β-D-glucopyranosyloxy]propoxy}-2-methyl-1,3-dioxane-2-carboxylic acid (11a) , 1H-NMR (D2O) δ: 4.56 , 4.44 , 4.16 , 4.02–3.99 , 3.95 , 3.06 , 3.82 , 3.40–3.29 , 2.00 , 1.00 ; ESI HRMS: m/z: calcd for C23H39NO17: 624.211; found: 624.2104 [M+ Na]+. This intermediate can be recycled.12 ; 1H-NMR (D2O) δ: 4.65 , 4.56–4.50 , 4.42 , 4.24–4.16 , 4.00–3.88 , 3.86–3.50 , 3.30–3.24 , 2.90 , 1.52 ; ESI HRMS: m/z: calcd for C25H38NO18: 640.2094; found: 640.2091 [M − H]−.The second fraction was the target compound R,S)-5-{3-(prop-2-yn)oxycarbonylamino-2-[α-D-galactopyranosyl-(1-4)-β-D-galactopyranosyl-(1-4)-β-D-glucopyranosyloxy]propoxy}-2-methyl-1,3-dioxane-2-carboxylic acid (13). was dissolved in water (1.5 mL) and neutralized with dry NaHCO3. Then other components were added in the following order: 1 M HEPES buffer , DTT solution , alkaline phosphatase , UDP-glucose  and crude fusion enzyme GalT/UDP-4'-epimerase +.pimerase  (0.3 mL)4-Nitrophenyl 2-(2-(2-(prop-2-ynyloxy)ethoxy)ethoxy)ethyl carbonate (15). To a solution of 2-(2-(2-(prop-2-ynyloxy)ethoxy)ethoxy)ethanol 14 +.arbonate . To a soR,S)-5-{2-[-β-D-glucopyranosyloxy]-5-oxo-6,9,12,15-tetraoxa-4-azaoctadec-17-ynyloxy}-2-methyl-1,3-dioxane-2-carboxylic acid (16). To a solution of 10  in methanol (12 mL) in a plastic tube NaOH  was added and the mixture was stirred at 80 °C for 3 days. The mixture was diluted with water and treated with dry ice until the solution pH was ~11 at which point a solution of 4-nitrophenylcarbonate 15  in methanol (2 mL) was added. The mixture was stirred overnight at room temperature. The reaction was not complete. Et3N  and more 15  were added and the mixture was stirred overnight. The reaction was not complete. The mixture was concentrated and purified by HPLC (C-18) using water with 0.1% of TFA and acetonitrile gradient to provide compound 16  as a white foam; 1H-NMR (D2O) δ: 4.55–4.54 , 4.43 , 4.23 , 4.22–4.15 , 3.98–3.92 , 3.91 , 3.81–3.50 , 3.37–3.26 , 2.89 , 1.50 ; ESI HRMS: m/z: calcd for C31H51NO21Na: 796.2846; found: 796.2841 [M+ Na]+.R,S)-5-{2-[-β-D-galactopyranosyl-(1-4)-β-D-glucopyranosyloxy]-5-oxo-6,9,12,15-tetraoxa-4-azaoctadec-17-ynyloxy}-2-methyl-1,3-dioxane-2-carboxylic acid (17). Lactose derivative 16  was dissolved in water (3 mL) and neutralized with dry NaHCO3. Then other components were added in the following order: 1 M HEPES buffer , DTT solution , alkaline phosphatase , UDP-glucose  and crude fusion enzyme GalT/UDP-4'-epimerase +.pimerase  (0.5 mL)Ntert-Butyloxycarbonylamino)propyl]-+): 265.15226; found: 265.15212.tert-Butyl 2-(2-((4-nitrophenoxy)carbonyloxy)ethoxy)ethylcarbamate (21). 2-(2-Aminoethoxy)-ethanol  was dissolved in methanol (10 mL) and di(tert-butoxy)carbonyl oxide  was added followed by triethylamine . After 30 min the mixture was concentrated and dried overnight under a vacuum provided by an oil pump. The residue was dissolved in dry DCM (20 mL) and p-nitrophenyl chloroformate  was added followed by pyridine . After 30 min the reaction was quenched by addition of methanol, then the mixture was diluted with DCM, washed with brine, concentrated and the residue was chromatographed on silica gel using hexane-acetone (4:1) to provide the product as slightly yellow syrup ; 1H-NMR (CDCl3) δ: 8.24 and 7.40 , 4.90 , 4.43 , 3.77 , 3.58 , 3.38–3.00 , 1.45 . ESI HRMS: m/z: calcd for C16H22N2O8Na ([M + Na]+): 393.12684; found 393.12652.N-(2-hydroxypropyl)methacrylamide with methacrylamide 19 (20).Copolymer of  A solution of N-(2-hydroxypropyl)methacrylamide  and 19  in degassed water (7 mL) was ultrasonicated under vacuum. A solution of ammonium persulfate (7 mg in 70 μL of water) was added to the mixture followed by an aq. solution of cysteamine hydrochloride . The reaction mixture was vortexed and incubated at 50 °C overnight then concentrated and dissolved in treated with TFA (8 mL). After 2 h the mixture was concentrated, the residue was dissolved in water, dialyzed extensively against deionized water, then freeze-dried to provide a white powder of aminated HPMA polymer 20 ; 1H-NMR (D2O) δ: 4.00–3.90 , 3.30–3.00 , 2.00–1.60 , 1.60–0.80 . Integration of 1H-NMR signals indicated ~5% incorporation of propylenediamine. 20 (70 mg) in dry DMF (1 mL) the linker 21  was added followed by triethylamine . The mixture was stirred overnight then diluted with water, dialyzed against deionized water and freeze-dried. The dry product was dissolved in TFA (2 mL) and stirred at room temperature for 30 min. The mixture was diluted with water (8 mL) carefully neutralized with dry sodium carbonate (1.33 g) in small portions. Then the solution was dialyzed against deionized water and lyophilized to give polymer 22 as white powder (62 mg); 1H-NMR (D2O) δ: 4.5 , 3.90 , 3.80 , 3.30–2.95 , 2.20–0.60 . To a solution of the polymer 22  was treated with linker 21 again as described for the synthesis of 22 to yield polymer 23 as white foam (78 mg); 1H-NMR (D2O) δ: 4.60–4.40 , 3.90 , 3.35–3.20 , 3.65 , 3.40–3.00 , 2.10–0.60 .Polymer 20  and activated carbonate 2  +): 265.1271; found 265.1271.hanamine   was performed at 70 °C in the presence of 4-cyanopentanoic acid dithiobenzoate (CTP) and 4,4'-azobis (ACVA) as chain transfer agent and initiator, respectively. In a typical protocol, AzMA  and HPMA  was dissolved in double distilled water containing a few drops of HCl (8 mL) followed by the addition of CTP  and ACVA  in 2-propanol (2mL) . The mixture was purged with nitrogen for 45 minutes and flask was placed in oil bath for polymerization under nitrogen for 24 h. The polymerization was quenched using liquid nitrogen and polymer was precipitated in acetone. The molecular weight and molecular weight distributions of copolymers were determined using gel permeation chromatography in aqueous media.The statistical copolymerization 5, 13 or 17, 1.5–2.0 eq) were dissolved in water (1 mL) and neutralized with dry NaHCO3 (to pH: 8), then 1 M sodium ascorbate (25 μL) and 0.05 M (50 μL) were added and the solution was left on a tumbler for 2 days. The reaction was checked by IR for the presence of azide. When no azido group could be detected the reaction mixture was diluted with water and extensively dialyzed against deionized water with the addition of EDTA, then filtered and lyophilized. Product was obtained in the form of white powder.An appropriate HPMA-AzMa copolymer (20–25 mg) and monomeric heterobifunctional ligand δ: 8.10 , 7.80–7.50 , 5.62 , 5.20 , 4.94 , 4.60–4.53 , 4.52 , 4.42-4.35 , 4.25–4.20 , 4.19–4.16 , 4.05–4.00 , 3.96–3.56 , 3.40–3.00 , 2.00–0.60 .HPMA-44/5-A; 1H-NMR (D2O) δ: 8.1 , 5.62 , 5.20 , 4.94 , 4.60–4.54 , 4.51 , 4.42 , 4.22 , 4.17 , 4.06–3.99 , 3.97–3.86 , 3.86–3.54 , 3.70–3.00 , 2.00–0.60 .HPMA-35/10-A; 1H-NMR (D2O) δ: 8.06 , 5.62 , 5.20 , 4.94 , 4.60–4.53 , 4.51 , 4.42–4.34 , 4.22–4.20 , 4.19–4.16 , 4.04–4.00 , 3.96–3.86 , 3.86–3.54 , 3.90–2.90 , 2.00–0.60 .HPMA-20/15-A; 1H-NMR (D2O) δ: 8.10 , 5.62 , 5.20 , 4.95 , 4.60–4.53 , 4.52 , 4.42–4.36 , 4.26–4.20 , 4.19–4.15 , 4.05–4.01 , 3.96–3.86 , 3.86–3.54 , 3.90–3.00 , 2.20–0.60 .HPMA-37/15-A; 1H-NMR (D2O) δ: 8.10 , 5.62 , 5.20 , 4.94 , 4.60–4.54 , 4.51 , 4.42–4.34 , 4.24–4.19 , 4.19–4.16 , 4.04–4.01 , 3.96–3.86 , 3.86–3.54 , 3.90–3.00 , 2.20–0.60 .HPMA-44/5-B; 1H-NMR (D2O) δ: 8.10 , 5.20 , 4.94 , 4.62–4.47 , 4.34 , 4.12–3.99 , 3.99–3.46 , 3.35–2.96 , 2.00–0.06 .HPMA-35/10-B; 1H-NMR (D2O) δ: 8.10 , 5.20 , 4.94 , 4.63–4.47 , 4.34 , 4.12–4.00 , 3.98–3.46 , 3.35–2.96 , 2.00–0.06 .HPMA-44/5-C; 1H-NMR (D2O) δ: 8.10 , 7.90–7.40 , 4.94 , 4.59–4.48 , 4.34 , 4.23–4.16 , 4.14–4.06 , 4.04–4.00 , 3.98–3.49 , 3.47–3.00 , 2.10–0.60 .HPMA-35/10-C; 1H-NMR (D2O) δ: 8.10 , 7.90–7.40 , 4.94 , 4.60–4.48 , 4.34 , 4.25–4.17 , 4.14–4.06 , 4.04–4.00 , 4.00–3.48 , 3.48–3.00 , 2.10–0.60 .n 977, Targeted molecular weight 70 kDa). The mixture was purged with nitrogen for 45 minutes and flask was placed in oil bath for polymerization under nitrogen for 24 h. The polymerization reaction mixture was quenched using liquid nitrogen and polymer was precipitated in acetone. The molecular weight and molecular weight distributions of copolymers were determined using aqueous gel permeation chromatography. The statistical copolymerization of 3-aminopropyl methacrylamide (APMA) and acrylamide (AA) was performed at 70 °C in the presence of 4-cyanopentanoic acid dithiobenzoate (CTP) and 4,4'-azobis (ACVA) as chain transfer agent and initiator, respectively. In a typical protocol AA  and APMA  was dissolved in dimethylsulfoxide (DMSO) (10 mL) followed by the addition of CTP  and ACVA  (targeted DPPAA-27/5. Poly[acrylamide-co-(3-aminopropylmethacrylamide hydrochloride)]  was dissolved in water (2.5 mL) and imidazole-1-sulfonyl azide  as described for preparation of PAA-27/5  as described for PAA-27/5. The title product obtained as off white powder (156 mg). IR: 2103.9 cm−1. 1H-NMR (D2O) δ: 3.44–3.34 , 3.30–3.12 , 2.38–1.98 , 1.82–1.34 , 1.2–1.08 .PAA-27/5-A. Azide PAA-27/5  and 5  were dissolved in water (1 mL) and pH adjusted to 8. Then 1 M sodium ascorbate (25 μL) and 0.05 M copper sulfate (50 μL). The reaction mixture was left on a tumbler for 2 days. Then it was dialyzed, filtered and freeze-dried to afford the polymer PAA-27/5-A as a white powder (35 mg). 1H-NMR (D2O) δ: 8.06 , 5.62 , 5.18 , 4.94 , 4.51 , 4.50–4.35 , 4.23 , 4.18 , 4.04–4.01 , 3.94–3.89 , 3.85–3.60 , 3.34–3.29 , 3.20–3.02 , 2.40–2.00 , 1.80–1.30 , 1.20–1.00 .PAA-72/5-A. Azide PAA-72/5  and 5  were dissolved in water (1 mL) and reaction and work-up were performed as described for PAA-27/5-A. The tile PAA-72/5-A was obtained as a white powder (35.3 mg). 1H-NMR (D2O) δ: 8.10 , 5.62 , 5.18 , 4.94 , 4.51 , 4.49–4.34 , 4.23 , 4.18 , 4.05–4.00 , 3.94–3.89 , 3.85–3.60 , 3.34–3.28 , 3.20–3.02 , 2.40–1.90 , 1.80–1.30 , 1.20–1.00 .n = 4–6 animals per group) were intravenously injected with a lethal dose(LD100 20 ng/g of body weight) of Stx1 that was premixed in a total volume of 100 μL with a heterobifunctional inhibitor in physiological saline solution. Forty-eight h after challenge, the mice were monitored every 4 h and immediately euthanized by CO2asphyxia if signs of Shigatoxemia (lethargy) became apparent. On the 10th day, all surviving mice were euthanized.The solid-phase binding-inhibition and Vero cell cytotoxicity-neutralization assays are fully described in previous reports ,18,19. Ip-nitrophenylcarbonate derivative of lactose 2 [1 provided protected lactose intermediate 3 with a linker armed with a terminal alkyne residue. After deprotection and subsequent enzymatic glycosylation of 4 the heterobifuntional ligand 5 was obtained  methacrylamide  and N-(3-tert-butoxycarbonylaminoxypropyl) methacrylamide (19) with cysteamine hydrochloride as a radical initiator. After deprotection of the product of polymerization, the HPMA polymer 20 randomly decorated with amino groups reacted with linker 21 to provide after deprotection polymer 22 with elongated side chains. The procedure was repeated to afford polymer 23 with twice elongated side chains . The RAFT copolymerization of HPMA with azide containing methacrylamide monomer (AzMA) was performed in aqueous conditions using 4,4'-azobis as the initiator and 4-cyanopentanoic acid 4-dithiobenzoate as the chain transfer agent , Table 1yl azide . 5, 13 and 17 to the azide-presenting scaffolds was accomplished via an efficient click reaction providing an array of polymeric heterobifunctional ligands: HPMA-16/5-A, HPMA-44/5-A, HPMA-35/10-A, HPMA-20/15-A, HPMA-37/15-A, HPMA-44/5-B, HPMA-35/10-B, HPMA-44/5-C, and HPMA-35/10-C, PAA-27/5-A, PAA-72/5-A.The conjugation of the heterobifunctional ligands Supramolecular inhibition is a novel strategy for the design of antagonists for pathogenic agents. The approach takes advantage of an endogenous protein such as an antibody or serum proteins of the innate immune system that serve as a template for presentation of a binding moiety specific to the target protein or cell ,24,25,26E. coli two main issues have to be resolved, namely, replacing polyacrylamide with a more biocompatible polymer, and controlling the molecular weight of the polymer-carrier to be below the kidney excretion cut-off. As the polyacrylamide backbone replacement we chose N-(2-hydroxypropyl)methacryamide (HPMA). Since its original use as a blood plasma expander [In order to address the feasibility of using polymeric heterobifunctional inhibitors such as PolyBAIT as injectable drugs to treat patients infected with enterohemorrhagic expander ,28. HPMAexpander . HPMA-n0, HPMA-n1 and HPMA-n2. These differed only in the length of the spacer-arm and were identical in all other respects since all of them were prepared from the same batch of aminated polyHPMA by sequential elongation of the linker. Inhibition data for both solid phase assay and cell intoxication assay presented in HPMA-n2 was still an order of magnitude lower than that of original polyacrylamide-based PolyBAIT . At the concentration, which was sufficient for the original PolyBAIT preparation to completely protect mice from the effects of Stx1 (200 μg/mouse), administration of HPMA-n2 resulted in 10–20 h delay of symptom onset, while complete protection was achieved at a 3 fold higher dose . This result confirmed the human SAP-dependant protective properties of tailored heterobifunctional polymers and the feasibility of transition of the methodology to a different polymer-carrier. Influences of length of the linker on the activity of the heterobifunctional polymer both in ELISA and cell culture assays suggest greater interference of the scaffold in HPMA polymers with formation of a supramolecular complex as compared with less bulky side chains in polyacrylamide. However, the question of how the nature of the polymer-carrier and its variable parameters such as molecular weight and payload influence the activity remained to be addressed.Conjugate her dose C. Again,in vivo and clinical testing of polymeric therapeutical and diagnostic agents. In the present work, we took advantage of the highly efficient “click reaction”, which enables conjugation of the active component to a polymer in aqueous media. The azido group of the AzMA monomer incorporated into HPMA backbone served as a reactive functionality, which permited nearly quantitative conjugation of carbohydrate ligands. Reversible addition–fragmentation chain transfer (RAFT) polymerization have previously been successfully employed to prepare HPMA copolymers with polydispersity of 1.1–1.8 (depending on molecular weight) containing methacrylamide monomers with a reactive group or cargo drug ,31,32. SHPMA-16/5-A with the lowest molecular weight, all compounds of the HPMA series have shown very similar activities regardless of molecular weight and also regardless of the payload (percent incorporation) of the binding fragments. Furthermore, in the case of ligands containing spatially separated binding fragments in each pendant grouping , only binding to site 2 provides the correct presentation of the pyruvate moiety to SAP. Site 3 is located too close the center pore and a heterobifunctional ligand would be unable to engage both proteins simultaneously at more than one or two points. Correct presentation of Pk-trisaccharide as a part of Gb3 glycolipid has recently been shown to play an important role in the interaction of Stx1 with the cell surface [The crystal structure of Stx1 in complex with Pccharide  revealedaffinity . Glucose surface . Hence,  surface . The optin vivo. We demonstrate that the inhibition is moderately influenced by the structure of the pendant ligand and the length of the spacer but is practically independent of molecular weight and the polymer density of the glycans. Our results suggest that steric bulk of the side chains in the polymeric scaffold impedes assembly of a supramolecular complex with endogenous serum protein SAP and requires heterobifunctional ligands that allow for greater separation between specific binding fragments. In summary, we have synthesized and evaluated a series of HPMA-based PolyBAITs, polymeric heterobifunctional ligands that inhibit Stx1 in a SAP-dependent manner. We observed that the inhibitory power of PolyBAITs in a solid-phase assay correlates with the cytotoxicity assay and efficacy


AbstractTorrenticola projectura and Hygrobates taiwanicus are described as new for science.New records of water mites (Acari: Hydrachnidia) from Baishih River drainage of north Taiwan, are presented. Twelve species are recorded, of which ten are new for Taiwan; two of them,  As the Tropic of Cancer bisects the Island, the climate changes from subtropical to tropical from the northern to southern parts of the island. The climate of Taiwan is PageBreakheavily influenced by monsoons and typhoons which bring an annual rainfall of ~ 2150 mm, with 80% of precipitation concentrated in the summer (or wet season), i.e. May–October. As there is a central mountain range extending from the north to the south of the island with surrounding plateau and hills, two third of the island is covered by mountainous area with an elevation 100–>3000 m. All rivers in Taiwan are characterized by short length and steep gradient with rapid flows and small drainage basins associated with the steep terrain. Such diverse patterns of geographical typology and hydrology result in the high levels of habitat heterogeneity and aquatic biodiversity of the Taiwan rivers . Water mites were sorted in the laboratory with the aid of a stereo microscope and preserved in 90% ethanol. All the material has been collected by Rita Yam and this is not repeated in the text. Holotypes and paratypes of the new species are deposited in the National Museum of Natural Science (NMNS), Taichung, Taiwan. Other materPageBreakials are kept in the collections of the Ecology and Conservation Laboratory, Department of Bioenvironmental Systems Engineering, National Taiwan University (ECL).In this study, water mites were collected at six sampling sites , 2A–F usIn the section ‘Material examined’, the sampling site codes were derived from the geographical database Rita Yam see . The comCook, 1967http://species-id.net/wiki/Hydrodroma_rheophilaECL-BA-1: 01.vii.2010 0/1/0 (mounted).PageBreakPageBreak specimen from the Chinkualiao stream shows general conformity with Hydrodroma rheophila. As Hydrodroma rheophila . ECL-BA-5: iii.2010 0/1/0.Sperchon rostratus Lundblad, 1969. The two females examined in the present study fit well the description of Burma, China (Guizhou Province), Turkey, Iran. New forTaiwan.Lundblad, 1941http://species-id.net/wiki/Sperchon_gracilipalpisECL-BA-2: iii.2010 1/1/0. ECL-BA-3: vi.2010 0/1/0 (mounted). ECL-BA-4: iii.2010 5/0 1/0/0 (mounted); iv.2010 0/1/0; 27.vii.2010 1/1/0. ECL-BA-5: iii.2010 1/1/0; iv.2010 0/1/0. ECL-BA-6: iv.2010 0/2/0.Sperchon gracilipalpis Lundblad, 1941. However, they resemble both Sperchon gracilipalpis and the Palaearctic Sperchon hispidus Koenike, 1895  muscle attachment plates; III/IV-L-3-5 with numerous pinnate dorsal setae; similar shape of palp . New forTaiwan.Lundblad, 1941http://species-id.net/wiki/Sperchon_cornutoidesECL-BA-1: 10.viii.2009 0/1/0; iii.2010 0/1/0; 01.vii.2010 0/1/0 (mounted); 02.vii.2010 0/2/0; 30.x.2010 1/0/0. ECL-BA-2: 27.vii.2010 2/0/0 (1/0/0 mounted). ECL-BA-3: 23.ix.2009 0/1/0; iii.2010 0/1/0. ECL-BA-4: iii.2010 0/1/0; 27.vii.2010 1/2/0 (1/1/0 mounted). ECL-BA-5: iv.2010 0/1/0. ECL-BA-6: iii.2010 0/1/0.Sperchon cornutoides Lundblad, 1941, a species known only from Java http://species-id.net/wiki/Monatractides_circuloidesPageBreak 17.vi.2009 3/0; iii.2010 2/9/0; vi.2010 0/1/0; 01.vii.2010 1/0/0; 27.vii.2010 1/0/0; 29.ix.2010 1/0.ECL-BA-1: 07.vii.2009 1/1/0; 23.ix.2009 2/0; iv.2010 13/0; 01.vi.2010 1/0; vii.2010 29/0; 27.vii.2010 2/1/0. ECL-BA-2: 02.ix.2009 11/2(1juvenile)/0; ii.2010 57/0; iii.2010 3/1/1; iv.2010 0/2/0; vi.2010 2/2/0; 27.vii.2010 17/0; 29.ix.2010 5/0; 30.x.2010 1/0. ECL-BA-3: 01.vii.2009 1/0/0; 23.vii.2009 1/0/0; ii.2010 4/11/0; ii.2010 10/0; iv.2010 0/1/0; 27.vii.2010 1/0/0. ECL-BA-4: 02.vii.2009 1/0/0; ii.2010 2/1/0; iii.2010 6/4/0; iv.2010 0/1/0; vi.2010 1/0/0; vii.2010 19/0; 27.vii.2010 5/0/0; 30.x.2010 1/0. ECL-BA-5: iii.2010 1/1/0; 01.vi.2010 4/0/0; vii.2010 11/0; 07.vii.2010 1/0/0; 30.x.2010 1/0/0. ECL-BA-6:Monatractides circuloides see: It is the most abundant species in our material. For an analysis of diagnostic characters of populations from Taiwan tentatively assigned to Malaysia, Thailand, Taiwan.Pešić, Semenchenko, Chatterjee, Yam & Chan, 2011http://species-id.net/wiki/Torrenticola_taiwanicusECL-BA-1: 07.vii.2009 1/0/0. ECL-BA-3: 27.vii.2010 1/4/0. ECL-BA-4: 02.vii.2009 0/2/0; ii.2010 0/1/0; iii. 2010 0/4/0; iv.2010 0/1/0 (mounted); vi.2010 1/1/0; 27.vii.2010 0/1/0; ECL-BA-5: 21.vii.2009 1/0/0; iii.2010 0/2/0; 01.vi.2010 1/1/0; 07.vii.2010 0/1/0. ECL-BA-6: ii.2010 1/0/0; iii. 2010 0/6/0; 27.vii.2010 0/1/0; 29.ix.2010 3/0.Torrenticola taiwanicus see: The studied material was collected from February to September. For an analysis of diagnostic characters of Taiwan.urn:lsid:zoobank.org:act:41E5931A-EFE2-42B6-B0D5-A86EB897B9FFhttp://species-id.net/wiki/Torrenticola_projecturaHolotype male (NMNS-6894-002), dissected and slide mounted, Taiwan, BA-1, Baishih River drainage, Chinkualiao Stream, 27.vii.2010. Paratype: one male, same data as holotype; one male, dissected and slide mounted, BA-2, Chinkualiao Stream, 02.ix.2009.Male . Cx-IV extending posterior to genital field; excretory pore posterior to line of primary sclerotization, Vgl-2 slightly posterior to excretory pore; capitulum with a short rostrum; medial suture line of Cx-II+III short; P-2 ventrodistal projection cone-shaped, pointed towards distal, P-3 ventrodistal projection larger than projection of P-2, P-3 with a long tapering ventral protrusion which curves distally.PageBreakratio 1.2 (1.2). Capitular bay L 150 (153), Cx-I total L 270 (263), Cx-I mL 119 (109), Cx-II+III mL 80 (70); ratio Cx-I L/Cx-II+III mL 3.4 (3.8); Cx-I mL/Cx-II+III mL 1.5 (1.6). Genital field L/W 156 (159)/124 (128), L/W ratio 1.26 (1.24); ejaculatory complex normal in shape, L 239 (211); distance genital field-excretory pore 180 (150), genital field-caudal idiosoma margin 272 (234). Capitulum (PageBreak segment/claw ratio 3.3 (3.3); palp  — Idiosoma ; palp  total L projectura’ - Latinised form of ‘projection’.The species is named after the distinctive shape of ventrodistal projection of P-3; ‘Torrenticola species.The shape of ventrodistal projection of P-3 of the new species is very distinctive and can separate it from all other Taiwan.urn:lsid:zoobank.org:act:8A67823A-EC83-418D-80B8-2934848B1D9Dhttp://species-id.net/wiki/Hygrobates_taiwanicusHolotype female (NMNS-6894-001), dissected and slide mounted, Taiwan, BA-3, Chinkualiao Stream, vi.2010. Paratypes: 0/1/0, same data as holotype; 1/1/0 (mounted), BA-1, Chinkualiao Stream, iii. 2010; 0/3/0, BA-4, Diyu Stream, iii.2010; 0/1/0, BA-5, Diyu Stream, iv.2010.Idiosoma L > 750; acetabula arranged in an obtuse angle; female pregenital sclerite with four setae, its anterior margin extending beyond anterior margin of genital plates; female genital plates longer than gonopore; posterior end of female genital plates posterior to postgenital sclerite; P-2 ventrodostal projection elongate, cone-shaped; proximoventral seta on P-4 longer and thicker than distoventral seta.PageBreakPageBreakdL of I-L-2-6 . Idiosoma L 775 (778), W 613 (706); integument soft, with very fine striation. Capitulum broadly fused with Cx-I. Coxae : coxal f I-L-2-6 : 69 (89) I-L-2-6 : 108 : similar to female, except for genital field. Idiosoma L 775-794, W 625-656; coxal field L 338, Cx-III W 463, Cx-I+II L/W 231/334; genital field L/W 160-170/148-151, anterior margin convex, L Ac-1-3: 51-58, 63-66, 63-66; ejaculatory complex L 169. Palp : total LThe species is named for its occurrence in Taiwan.Hygrobates species belong to the group of the species characterized byt the presence of two or three setae upon, and inside the margin of, the female pregenital sclerite and proximoventral seta on P-4 longer and thicker than distoventral seta. This group includes three species ; 29.ix.2010 0/1/0.Hygrobates species from the Baishih River drainage resemble to the Palaeractic Hygrobates longiporus Thor. In shape of coxal . ECL-BA-5, 20.viii.2009 0/1/0.Hygrobates hamatus. This species is very similar to Hygrobates soari K. Viets, 1911, a species widspread in the Afrotropical region, reaching in its distribution to northern Oman http://species-id.net/wiki/Atractides_spatiosusECL-BA-1: 01.vi.2009 0/1/0; 02.vii.2010 0/1/0. ECL-BA-2: iv.2010 0/2/0; vi.2010 0/1/0. ECL-BA-3: 03.ix.2009 0/3/0. ECL-BA-4: iii.2010 0/4/0 (0/1/0 mounted); iii.2010 0/1/0. ECL-BA-5: iv.2010 0/1/0; 07.vii.2010 0/4/0. ECL-BA-6: iii.2010 0/1/0; iv.2010 0/2/0; vi.2010 0/1/0; ix.2010 0/1/0.Atractides spatiosus . A problem exists regarding the similarity between Atractides spatiosus and Atractides cognatus , a further taxon originally introduced as a subspecies and later elevated to species rank by Pešić and Smit (2009). However, the populations attributed by Pešić and Smit (2009) to Atractides cognatus differ from the original description of the later (see: K. Atractides cognatus), and probably represented an undescribed species. For the time being, the sclerotization of the excretory pore http://species-id.net/wiki/Atractides_cognatusECL-BA-1: 23.ix.2009 0/1/0. ECL-BA-4: 20.viii.2009 0/1/0; 27.vii.2010 0/2/0. ECL-BA-5: 27.vii.2010 0/3/0; x.2010 0/1/0. ECL-BA-6: 01.vii.2010 0/1/0; 27.vii.2010 0/1/0; 30.x.2010 0/1/0 (mounted).Atractides cf. spatiosus.See discussion under PageBreakPageBreakSE Asia. New for Taiwan.


The structures were determined by extensive spectroscopic analyses and by comparison with the spectral data of related known compounds. Metabolite 1 is rarely found in spatane skeletons reported from soft corals. In addition, compound 1 exhibited weak cytotoxicity towards human tumor cell lines T-47 D and K-562.A new spatane diterpenoid, leptoclalin A ( Klyxum simplex +), implying five degrees of unsaturation. The IR spectrum of 1 revealed the presence of a hydroxy functionality (νmax = 3365 cm−1). The 13C-NMR and DEPT spectroscopic data  showed the presence of a hydroxy group. A 1,1-disubstituted and a 1,2-disubstituted double bond were also identified from NMR signals appearing at δC 157.5 (C), 103.7 (CH2), and δH 4.74  and 4.70 , and at δC 139.2 (CH), 125.8 (CH), and δH 5.58 , respectively. Moreover, the 1H NMR spectroscopic data of 1 revealed evidence of four methyl groups at δ 0.83 , 1.00  and 1.31 .Leptoclalin A , it was possible to establish the proton sequences from H2-2 to H-10 through H2-3; H2-5 to H2-6; H-8 to H-10 through H-9; and H-13 to H3-14. The overlapping of proton singals of H-7 and H-8 at δH 1.62 ppm, measured in CDCl3, was clearly resolved by measuring the 1H NMR spectrum in pyridine-d5  were observed between H-6 to H-13 through H-7. Therefore, the 1H–1H COSY experiment allowed the building of the two partial structures of the consecutive proton spin systems indicated in bold in 2-2 to C-1, H2-3 to C-1 and C-9, H2-5 to C-4 and C-10, H2-6 to C-4 and C-8, H-9 to C-7 and H-10 to C-8 established the connectivity within the 5-membered, 4-membered and 5-membered rings. Moreover, the correlations of H2-11 to C-2 and C-9 indicated the attachment of an sp2 methylene at C-1, and the methyl was attached at C-4 on the basis of the correlations of H3-12 to C-4, C-5, C-8 and C-10. The planar structure of the side chain was elucidated mainly from the key HMBC correlations from H3-14 to C-7, C-13, and C-15, H2-15 to C-16 and C-17, and both methyls H3-19 and H3-20 to C-18 and C-17, and thus the connectivity from C-13 to C-20 was fully established. The above findings suggested that 1 has a tricyclic carbon skeleton. The planar structure and all of the eriments . From the-d5 see  into two1 , it was found that H-9 (δ 2.46) showed NOE interactions with H-7 (δ 1.62) and H-10 (δ 2.24); therefore, assuming the β-orientation of H-9, H-7 and H-10 should also be positioned on the β face. One of the methylene protons at C-3 (δ 1.52) exhibited NOE correlations with H-10 and was characterized as H-3β, while the other (δ 1.74) was designated H-3α. NOE correlations observed between H-3α and H3-12 (δ 0.94) and H3-12 and H-8 (δ 1.67) reflected the α-orientations of H3-12 and H-8. Moreover, NOE correlations were observed between H-8 with H-13 but not with H3-14, and between H-7 with H3-14, indicating that H3-14 has a β-orientation. Furthermore, the configuration of the double bond at C-16/C-17 was determined by comparison of the NMR data of 1 in CDCl3 with those of two related synthetic compounds, (23E)-cycloart-23-ene-3β,25-diol (4) and (23E)-cycloart-23-ene-3β,25-diol (5) . Thus, it was suggested that the geometry of 1 at C-16/C-17 is E. On the basis of the above analysis, the structure of 1 was established.A computer-modeled 3D structure of 1  was genediol (5) , also mel (5) , HCT 116 , T-47D (hormone-dependent breast cancer), and K-562 (human chronic myelogenous leukemia). Cells were treated with different concentrations of 1–3 for 72 h. The results show that compound 1 was found to show weak cytotoxicity towards the growth of T-47D and K-562 tumor cells  (50 > 20 μg/mL) towards the above four cancer cell lines -2,5-diphenyl tetrazolium bromide (MTT) assay to examine the cytotoxic activities of compounds ctively) . The othll lines .1H-NMR and 100 MHz (or 125 MHz) for 13C-NMR, respectively, in CDCl3 and pyridine-d5. ESI-MS-spectra were obtained with a Bruker APEX II mass spectrometer. Gravity column chromatography was performed on silica gel . TLC was carried out on precoated Kieselgel 60 F254  and spots were visualized by spraying with 10% H2SO4 solution followed by heating. High-performance liquid chromatography (HPLC) was performed using a system comprised of a Hitachi L-7100 pump and a Rheodyne 7725 injection port. A preparative normal phase column  was used for HPLC.Optical rotation values were measured with a Jasco P-1010 digital polarimeter. IR spectra were recorded on a Varian Digilab FTS 1000 Fourier transform infrared spectrophotometer. The NMR spectra were recorded on a Varian Mercury Plus 400 FT-NMR (or Varian Unity INOVA 500 FT-NMR) instrument at 400 MHz (or 500 MHz) for Sinularia leptoclados were collected off the coast of Pingtung, southern Taiwan, and transplanted to a 120-ton cultivating tank equipped with a flow-through sea water system in June 2005. The cultured soft coral was harvested in December 2010. A voucher specimen (specimen No. 2010CSC-2) was deposited in the National Museum of Marine Biology and Aquarium, Pingtung, Taiwan.Specimens of the soft coral Sinularia leptoclados (12.5 kg fresh wt) was frozen for storage and then freeze dried. The freeze-dried material (3.0 kg) was minced and extracted exhaustively with EtOAc (5 × 5 L). The EtOAc extract was evaporated to yield a residue (100.5 g), which was subjected to open column chromatography on silica gel, eluting with n-hexane (H)–EtOAc (E) gradient and EtOAc (E)–acetone (A) gradient to give 12 fractions: Fr-1 (eluted by H–E 100:1), Fr-2 (eluted by H–E 50:1), Fr-3 (eluted by H–E 30:1), Fr-4 (eluted by H–E 20:1), Fr-5 (eluted by H–E 10:1), Fr-6 (eluted by H–E 8:1), Fr-7 (eluted by H–E 5:1), Fr-8 (eluted by H–E 3:1), Fr-9 (eluted by H–E 1:1), Fr-10 (eluted by EtOAc), Fr-11 (eluted by E–A 1:1) and Fr-12 (eluted by acetone). Fraction 5 (2.2 g) was separated by silica gel column chromatography with gradient elution  to yield five subfractions (5A–5E). Subfraction 5B (108 mg) was subjected to normal-phase HPLC with n-hexane–EtOAc (10:1) elution to afford 1 (3.4 mg). Fraction 8 (6.5 g), was further separated by silica gel open column chromatography with gradient elution  to yield seven subfractions (8A–8G). Subfraction 8C (4.5 g) was further chromatographed over silica gel using n-hexane–acetone (5:1) to afford 3 (750 mg) and a mixture, which was further purified by normal phase HPLC using n-hexane–EtOAc (3:2) to afford 3 (350 mg) and 2 (1.2 g), respectively.The soft coral 1): colorless oil; [α]24D +24 ; IR (neat) νmax 3365, 2927, 2865, 1456, and 1374 cm−1; 13C and 1H NMR data, see m/z 311 [M + Na]+; HRESIMS m/z 311.2353 [M + Na]+ .Leptoclalin A -2,5-diphenyltetrazolium bromide] colorimetric method [Cell lines were purchased from the American Type Culture Collection (ATCC). Cytotoxicity assays of compounds c method ,18. Spatoglossum schmittii from the Galapagos Islands [Sinularia leptoclados led to the obtainment of one new spatane diterpenoid (1). Moreover, to the best of our knowledge, metabolite 1has a spatane skeleton rarely found in soft corals.Spatane diterpenoids were first reported in 1980 as having been obtained from the brown alga  Islands ,25,26,27 Islands . Our inv


Villosiclava virens of rice false smut. Ustiloxins A and B as two main mycotoxins were determined conveniently by LC-ESI-MS in the water extract from rice false smut balls which were mostly composed of the chlamydospores and mycelia of the pathogen. Both ustiloxins A and B in the water extract were also quantitatively analyzed by HPLC. This is the first report on the determination and analysis of ustiloxins A and B simultaneously by LC-ESI-MS and HPLC in false smut balls of rice.Ustiloxins are cyclopeptide mycotoxins produced by the pathogenic fungus  Villosiclava virens (Nakata) Tanaka & Tanaka (anamorph: Ustilaginoidea virens Takahashi) + . 1H NMR  δ (ppm): 7.60 , 7.08 , 4.93 , 4.84 , 4.35–4.40 , 4.28 , 4.14 , 3.99 , 3.77 , 3.33 , 3.04 , 2.77 , 2.17–2.25 , 2.12 , 1.86–1.92 , 1.76 , 1.68–1.73 , 1.09 , 0.88 , 0.78 . 13C NMR (D2O) δ (ppm): 176.0 (C-20), 174.1 (6′-C), 170.7 (C-5), 170.0 (C-17), 166.0 (C-8), 151.9 (C-14), 145.7 (C-15), 136.1 (C-12), 127.7 (C-11), 123.9 (C-16), 113.7 (C-13), 86.9 (C-2), 73.7 (C-10), 66.4 (C-9), 64.5 (2′-C), 63.5 (3′-C), 59.8 (6-C), 59.3 (3-C), 52.5 (5′-C), 43.5 (19-C), 36.4 (4′-C), 32.0 (NCH3-C), 31.8 (22-C), 28.4 (24-C), 20.9 (21-C), 18.0 (26-C), 17.7 (25-C), 7.5 (23-C). The data were consistent with the literature + . 1H NMR  δ (ppm): 7.57 , 7.39 , 4.98 , 4.71 , 4.46 , 4.33–4.38 , 4.21 , 3.97 , 3.81 , 3.75 , 3.37 , 3.03 , 2.76 , 2.04–2.19 , 1.74 , 1.65–1.70 , 1.19 , 0.97 . 13C NMR (D2O) δ (ppm): 176.2 (C-20), 174.1(6′-C), 171.8 (C-5), 169.8 (C-17), 165.6 (C-8), 152.0 (C-14), 145.6 (C-15), 136.5 (C-12), 127.8 (C-11), 123.9 (C-16), 113.7 (C-13), 87.0 (C-2), 73.3 (C-10), 66.1 (C-9), 64.5 (2′-C), 63.5 (3′-C), 59.6 (3-C), 52.4 (5′-C), 49.4 (6-C), 43.5 (19-C), 36.4 (4′-C), 31.7 (NCH3-C), 30.9 (22-C), 21.6 (21-C), 15.2 (24-C), 7.7 (23-C). The data were consistent with the literature [2 was identified as ustiloxin B.Compound v/v) as the mother solution at 1000 μg/mL for each ustiloxin. The mother solution was further diluted into 150 μg/mL, 125 μg/mL, 100 μg/mL, 62.5 μg/mL, 50 μg/mL, 31.25 μg/mL, 25 μg/mL and 12.5 μg/mL samples with methanol-water , and the solutions were kept at 4 °C.An amount of 1 mg of ustiloxin A or ustiloxin B was accurately weighed. They were mixed and dissolved in 1 mL of methanol-water . It was then filtered through a filter  before analysis.Based on the physicochemical properties of the ustiloxins, each powdered sample of rice false smut balls (200 mg) was extracted with deionized water (3 × 6 mL) in an ultrasonic bath at room temperature. The extraction time for each extraction was 4 h. The water extract was then concentrated under vacuum at 60 °C by a rotary evaporator to dryness and dissolved in 5 mL of methanol-water  for the validation of analytical methods ,13.n = 5). Similarly, the inter-day precision was assessed by analyzing the aforementioned three levels of ustiloxins A and B on three separate days (n = 9). The peak areas of the ustiloxins A and B and the relative standard deviations (RSDs) were calculated [The intra-day precision was examined by analysis of authentic ustiloxins A and B at levels 150 μg/mL, 50 μg/mL and 12.5 μg/mL, which were prepared and analyzed on the same day. Each level of the ustiloxins A and B was detected for five repetitions  in which the contents of ustiloxins A and B were known. The added amounts of authentic ustiloxins A and B were at low, medium, and high levels. The samples were prepared in triplicate, and the recovery percentages and RSDs were calculated.The limit of detection (LOD) and limit of quantification (LOQ) were based on signal-to-noise approach. Determination of the signal-to-noise ratio is performed by comparing measured signals from samples with known low concentrations of analyte with those of blank samples and by establishing the minimum concentration at which the analyte can be reliably detected or quantified . A signaThe contents of ustiloxins A and B in rice false smut balls were determined by HPLC-UV. The solutions of the crude extracts were prepared as described in Section 3.5.2. The data were presented as an average of six replicates for each sample. The experiments were repeated three times. The relative standard deviations (RSDs) were also calculated.i.e., ustiloxins C and D) from rice false smut balls are in progress. It is worth to mention that ustiloxins have been considered as the antimitotic compounds that may exhibit a broad range of biological activities and can be used for medicinal and agrochemical purposes, acting as anticancer, antifungal, and anthelmintic agents [In this study ustiloxins A and B as the two main mycotoxins were determined conveniently by LC-ESI-MS in the water extract from rice false smut balls. Both ustiloxins A and B in rice false smut balls were also quantitatively analyzed by HPLC-UV. To the best of our knowledge, this is the first report on the determination and analysis of ustiloxins A and B in rice false smut balls by LC-ESI-MS and HPLC methods simultaneously. The LC-ESI-MS method should be a practical way for rapid determination and analysis of the main ustiloxins in test samples. Isolation and structural identification of other ustiloxins (c agents ,16. Morec agents –19.


The structures of the new compounds were elucidated on the basis of mass spectrometric and NMR spectroscopic analysis. Compounds 1–4 are derivatives of siphonodictyal E (9). Siphonodictyal E4 (4) exhibited mild antiproliferation activity against L929 mouse fibroblast, KB-31 epidermoid carcinoma, and MCF-7 breast cancer cell lines, while siphondictyal E3 (3) and cyclosiphonodictyol A (5) showed moderate activity against Gram-positive bacteria.Four new sulfated sesquiterpene hydroquinones siphonodictyals E1–E4 ( Aka coralliphagum  is known to have four distinct morphological forms: forma typica, f. tubulosa, f. obruta, and f. incrustans − which indicated the molecular formula C24H35NO11S2. Comparison of the NMR data of 1 and siphonodictyal E . Moreover, the 1H,1H-COSY correlation between the two methylene groups CH2-23 and CH2-24 as well as the 1H,13C-HMBC correlations from H-22 (δ 8.67) to C-18 (δ 144.0), C-19 (δ 156.4), C-20 (δ 110.9), C-21 (δ 141.8), and C-23 (δ 51.2) further indicated that an iminoethanesulfonic acid is connected to the aromatic ring system in 1  wgeometry –7. The Ntem in 1 , as in s in 1 6,  5]..1) was i2) revealed a high degree of similarity with the data reported for siphonodictyal E (9). However, the molecular ion peak of compound 2 was shown at m/z 469.1534 ([M − H]− C22H29O9S), which has 80 amu more than that of 9, indicating an additional sulfate ester group in compound 2. The 1H,13C-HMBC correlations from 18-OH to C-17 (δ 122.8), C-18 (δ 141.8), and C-19 (δ 147.8) as well as from 19-OH to C-18 (δ 141.8), C-19 (δ 147.8), and C-20 (δ 115.5) indicated that C-18 and C-19 are connected to hydroxy groups rather than sulfate ester groups  r3) displayed the pseudomolecular ion peak [M + Na − 2H]− at m/z 539.1046, indicating the molecular formula C22H30O10S2. The 13C NMR spectrum contained six olefinic signals in addition to the aromatic signals, and the 1H NMR spectrum contained four olefinic methyl signals at δ 1.62 (s), δ 1.54 (s), δ 1.55 (s), and δ 1.66 (s), which suggested a sesquiterpene side chain containing three trisubstituted olefinic bonds. The two sulfate ester groups were assigned to attach to C-18 and C-21 after comparison of the NMR data of the aromatic moieties of 3 with those of siphonodictyal B3  spectrum for siphonodictyal E3  showed the pseudomolecular ion peak [M + Na − 2H]− at m/z 555.0954, confirmed its molecular formula as C22H30O11S2, corresponding to eight double bond equivalents. The sesquiterpenoid moiety of the molecule contained two trisubstituted olefinic bonds  and a hemiacetal carbon (δ 96.2) at one terminus which differed from the acyclic side chain in 1 and 2. The remaining six carbon signals in the downfield region of the 13C NMR spectrum belonged to the tetrasubstituted aromatic ring system. In contrast to 1 and 2, compound 4 contained an oxymethine group  which connected the sesquiterpene chain with the aromatic ring. This is supported by the six/seven HMBC correlations from H-22 to C-1 (δ 96.2), C-2 (δ 37.3), C-3 (δ 31.4), C-13 (δ 19.9), C-19 (δ 143.2) / C-21 (δ 142.9), and C-20 (δ 120.3). The HMBC correlations from H-12  to C-17 (δ 120.8) and C-21 and from H-17 (δ 7.19) to C-18 (δ 137.2) confirmed the position of the unsubstituted CH-17 as shown in the proposed structure (4a and 4b (3-ox in 8) − at m/z 343.2292, confirmed its molecular formula as C22H32O3. The 13C NMR spectrum displayed six signals in the area of aromatic carbon atoms and 16 signals in the aliphatic region. The 1H,13C-HSQC spectrum indicated four methyl and seven methylene groups. Two of these methylene groups connected directly to the aromatic ring system indicated by the 1H,13C-HMBC correlations H-15  → C-16 (δ 129.9), C-17(δ 146.4), and C-21 (δ 127.7) as well as H-22  → C-16, C-20 (δ 146.5), and C-21. The sesquiterpene unit of 5 was identified by 1H,1H-COSY and 1H,13C-HMBC experiments. Furthermore, the HMBC correlations H-15 → C-8 (δ 78.5), C-9 (δ 58.1), and C-10 (δ 38.2) confirmed the connection between C-15 and C-9 and the HMBC correlation H-22 → C-8 indicated the existence of the oxygen bridge between C-22 and C-8. The p-hydroquinone moiety was established on the basis of the 1H and 13C NMR chemical shifts as well as the HMBC correlations mentioned above, namely, H-15 → C-16, C-17, C-21; H-22 → C-16, C-20, C-21. The relative configuration was confirmed by a ROESY experiment. The observed ROESY correlations between H-12 and H-13, H-13 and H-15, H-14 and H-15 suggested that the three methyl groups as well as CH2-15 are axial. Examination of the NMR data for 5 revealed a high degree of similarity with data reported for bis(sulfato)cyclosiphonodictyol A − calcd for C24H33NO11S2Na, 598.1398; found, 598.1338.Siphonodictyal E1 (2): yellow powder; 1H NMR  and 13C NMR  see m/z: [M − H]− calcd for C22H29O9S, 469.1538; found, 469.1561.Siphonodictyal E2 (3): yellow powder; 1H NMR  and 13C NMR  see m/z: [M + Na − 2H]− calcd for C22H28O10S2Na, 539.1027; found, 539.1046.Siphonodictyal E3 (4): yellow powder; CD data {(MeOH)  = +37}; 1H NMR  and 13C NMR  see m/z: [M + Na − 2H]− calcd for C22H28O11S2Na, 555.0976; found, 555.0954.Siphonodictyal E4 (5): colorless powder; CD data {(MeOH)  = +163}; 1H NMR  and 13C NMR  see m/z: [M − H]− calcd for C22H31O3, 343.2279; found, 343.2292.Cyclosiphonodictyol A (File 11D and 2D NMR spectra of the isolated compounds.


Background: 5-(Ethylsulfonyl)-2-methoxyaniline (5) is part of the structure in 131 compounds possessing different biological activities. In most cases, they have antitumor properties (112 compounds). Other compounds are described as cardiovascular agents, ion-channel blockers, nervous-system blockers, anti-inflammatory agents, or antidiabetic, antiosteoporotic and hypolipemic species. Compound 5 is a precursor of different protein-kinase inhibitors or enzyme modulators . The structure of 5 represents a fragment for several powerful inhibitors of VEGFR2, a key angiogenic receptor. Antiangiogenic inhibitors slow down or stop new blood-vessel formation from pre-existing vasculature. Some antiangiogenic drugs inhibiting the VEGFR2 receptor are successfully used in clinics for the treatment of several types of tumours in synergy with chemotherapy . The structure of 5 is an important pharmacophoric fragment of potent VEGFR2 inhibitors . Up to now, 25 VEGFR2 inhibitors possessing a fragment of 5 can be found in the literature. Despite the high significance of 5-(ethylsulfonyl)-2-methoxyaniline (5) its preparation has not yet been described.Results: Here we have developed a convenient synthesis of important polyheterosubstituted aniline 5 starting from commercially available 4-methoxybenzene-1-sulfonyl chloride (1) in four steps and 59% overall yield. The target 5-(ethylsulfonyl)-2-methoxyaniline (5) and its synthetic intermediates 2–4 together with a new compound 5-(ethylsulfonyl)-2-methoxy-1,3-dinitrobenzene (4a) have been precisely physicochemically characterised. Compound 5 is used for the development of small organic compounds, i.e., modulators targeting a broad spectrum of important human protein receptors or enzymes, e.g., VEGFR2, EGFR, PDGFR, TEK kinase, ckit, EphB4, ErbB-2 receptor tyrosine kinase, cyclin-dependent kinases 2 and 4, neu receptor, polo-like kinase 1, alpha and beta adrenoreceptors, glycogen phosphorylase, IMP dehydrogenase, MMPs 2, 3, 9 and 13, etc. oxazole-5-sulfonyl chloride (E) by reaction of D with (EtO)3CH in the presence of p-TsOH. Reduction of sulfonyl chloride E to sodium sulfinate F by Na2SO3 and Na2CO3 and its reaction with ethyl iodide proceeded as nonselective reactions, which excluded this methodology from further efforts . Compound 1 can be prepared easily from methoxybenzene (anisole) and ClSO3H in 88% yield , is a commercially available compound. This substance can be prepared in 88% yield from readily available anisole, sulfuryl chloride, H2SO4 and DMF according to the literature , is not a commercially available compound. Sulfinate 2 can be prepared from arylsulfonyl chloride 1 by treatment with Na2SO3, NaHCO3 at 65 °C in 98% yield , is not a commercially available compound from current suppliers. The Reaxys database contains different mp values for arylethylsulfone 3: 40–41°C [EtOH] [2O]  , 55–56 °OH] [2O]  , CAS 75-79-0, i5% yield . None of4-(Ethylsulfonyl)-1-methoxy-2-nitrobenzene (4), CAS [51572-44-2], is not commercially available from Sigma-Aldrich, Merck or Acros Organics. No direct synthesis or experimental physicochemical properties have been described for this compound in the SciFinder database. In the Reaxys database only one mp, i.e., 119–120 °C was present , is a compound with only limited commercial availability. For this compound no synthesis can be found in the Reaxys and SciFinder databases. There were 37 reactions in Reaxys and 191 reactions in SciFinder described. In all cases compound 5 was exclusively a starting material or reactant. Experimental IR and 1H NMR (CDCl3) spectra for compound 5 were mentioned in the SciFinder database and originated from Bio-Rad Laboratories data collection ; IR (neat) ν/cm−1: 3358 , 2959 (m), 2837 (m), 1591 (s), 1575 (m), 1490 (s), 1459 (m), 1441 (m), 1401 (w), 1299 (m), 1244 (s), 1188 (m), 1171 (m), 1082 (m), 1046 (m), 1029 (s), 974 (s), 835 (m), 824 (w), 815 (m), 793 (m), 722 (w); 1H NMR  δ 7.42  = 8.7 Hz, 2H, H-C(2)), 6.87  = 8.7 Hz, 2H, H-C(3)), 3.74 ; 13C NMR  δ 159.0 (C4-O), 127.0 (C1-S), 125.5 (C2), 112.8 (C3), 55.0 (MeO-); ESIMS m/z: 171.1 [M − Na]−; Anal. calcd for C7H7NaO3S (194.18): C, 43.30; H, 3.63; S, 16.51; found: C, 43.15; H, 3.48; S, 16.22%.Synthesis of 1-(ethylsulfonyl)-4-methoxybenzene (3). A crude product from the previous reaction 12.20 g  2 was dissolved in 150 mL of MeOH. The solution was stirred and heated under reflux in an Ar atmosphere. When the reaction mixture started to boil, 12.6 mL  EtI was added dropwise over 10 min via syringe and septum. The reaction mixture was heated under reflux and monitored by TLC. After 2 h, the mixture was left to cool to rt and subsequently evaporated by RVE to give a pale yellow solid material. The crude product 3 was triturated with CHCl3 (3 × 30 mL), the suspension filtered and the organic solution concentrated by RVE to 1/10 of its original volume. The solution crystallized by standing in a refrigerator overnight to give white crystals. The crystals were filtered off and washed with cold hexane to yield 11.30 g  of the product 3. Pale yellow solid; mp 54.9–56.4 °C [CHCl3] (lit. 55–56 °C [EtOH/H2O] +; Anal. calcd for C9H12O3S (200.25): C, 53.98; H, 6.04; S, 16.01; found: C, 53.88; H, 5.90; S, 16.42%.tOH/H2O] ); IR (neSynthesis of 4-(ethylsulfonyl)-1-methoxy-2-nitrobenzene (4). 1-(Ethylsulfonyl)-4-methoxybenzene (3) 11.30 g  was dissolved in 140 mL of HNO3 . The reaction mixture was heated to 100 °C and monitored by TLC. A complete conversion of starting material 4 was observed within 2 h. Then the mixture was left to cool to rt and carefully poured into 250 mL of ice-cold water. A white solid material precipitated. Then the suspension was washed with CHCl3 (3 × 60 mL). The combined organic phase was dried over Na2SO4 and filtered, and the solution was evaporated by RVE to yield 10.14 g  4 in the form of a pale yellow solid material. Yellow solid; mp 118.5–119.7 °C [CHCl3] (lit. 119–120 °C ; IR (neat) ν/cm−1: 3075 (m), 2974 (w), 1611 (m), 1536 , 1481 (m), 1457 (w), 1428 (w), 1406 (w), 1344 (m), 1314 (s), 1279 (m), 1260 (m), 1235 (w), 1211 (m), 1184 (m), 1130 (s), 1094 (m), 1047 (m), 968 (m), 927 (w), 912 (m), 890 (m), 785 (w), 772 (m), 737 (m), 711 (m), 686 (w); 1H NMR  δ 8.52 ), 4.15 , 3.23  = 7.4 Hz, 2H, SO2CH2), 1.38  = 7.4 Hz, 3H, SO2CH2CH3); 13C NMR  δ 151.5 (C2-O), 145.2 (C3), 134.7 (C5), 129.0 (C4), 65.3 (MeO), 50.8 (SO2CH2), 7.5 (SO2CH2CH3); ESIMS m/z: 290.1 [M]−; Anal. calcd for C9H10N2O7S (290.25): C, 37.24; H, 3.47; N, 9.65; S, 11.05; found: C, 36.89; H, 3.50; N, 9.15; S, 10.97%.Synthesis of 5-(ethylsulfonyl)-2-methoxyaniline (5). 4-(Ethylsulfonyl)-1-methoxy-2-nitrobenzene (4) 6.38 g  was dissolved in 150 mL of EtOH. A catalytic amount of 10% Pd/C  was added to the solution. The mixture was stirred under an atmosphere of H2 at 34 °C and regularly analysed by TLC. After 2 days the reaction was completed. The mixture was filtered through a small column filled with silica gel and the solution evaporated by RVE. The crude compound was dried by HV. Afterwards 5.01 g  of a solid powder was obtained as the pure product 5 according to the 1H NMR spectrum. For mp determination a crystallization of 5 from MeOH was performed. Solid; mp 102.9–103.8 °C [MeOH]; IR (neat) ν/cm−1: 3459 , 3366 , 3080 (w), 2966 (m), 2944 (m), 2924 (m), 2843 (m), 1616 (m), 1578 (m), 1504 (m), 1470 (m), 1458 (m), 1426 (m), 1375 (w), 1354 (w), 1303 (m), 1287 (s), 1277 (s), 1228 (s), 1190 (w), 1137 (s), 1108 (s), 1074 (m), 1046 (m), 1018 (s), 919 (m), 874 (m), 819 (m), 759 (m), 735 (m), 719 (s), 660 (m); 1H NMR  δ 7.25  = 8.4, J = 2.3 Hz, 1H, H-C(4)), 7.19  = 2.3 Hz, 1H, H-C(6)), 6.87  = 8.4 Hz, 1H, H-C(3)), 4.15 , 3.92 , 3.07  = 7.4 Hz, 2H, SO2CH2), 1.24  = 7.4 Hz, 3H, SO2CH2CH3); 13C NMR  δ 151.1 (C2-O), 137.2 (C1), 130.3 (C5), 119.3 (C4), 113.3 (C6), 109.9 (C3), 56.0 (MeO), 50.9 (SO2CH2), 7.8 (SO2CH2CH3); ESIMS m/z: 216 [M + H]+; Anal. calcd for C9H13NO3S (215.27): C, 50.21; H, 6.09; N, 6.51; S, 14.90; found: C, 50.50; H, 6.15; N, 6.75; S, 14.77%.


Sinularia sp., collected from Bowden Reef, Queensland, Australia, yielded ten natural products. These included the new nitrogenous diterpene -4-methoxy-9-((dimethylamino)-methyl)-12,15-epoxy-11(13)-en-decahydronaphthalen-16-ol (1), and the new lobane, -loba-8,10,13(14),15(16)-tetraen-17,18-diol-17-acetate (2). Also isolated were two known cembranes, sarcophytol-B and -11,12-epoxycembratrien-15-ol, and six known lobanes, loba-8,10,13(15)-triene-16,17,18-triol, 14,18-epoxyloba-8,10,13(15)-trien-17-ol, lobatrientriol, lobatrienolide, 14,17-epoxyloba-8,10,13(15)-trien-18-ol-18-acetate and (17R)-loba-8,10,13(15)-trien-17,18-diol. Structures of the new compounds were elucidated through interpretation of spectra obtained after extensive NMR and MS investigations and comparison with literature values. The tumour cell growth inhibition potential of 1 and 2 along with loba-8,10,13(15)-triene-16,17,18-triol, 14,17-epoxyloba-8,10,13(15)-trien-18-ol-18-acetate, lobatrienolide, -11,12-epoxycembratrien-15-ol and sarcophytol-B were assessed against three human tumour cell lines . The lobanes and cembranes tested demonstrated 50% growth inhibition in the range 6.8–18.5 µM, with no selectivity, whilst 1 was less active (GI50 70–175 µM).The methanol extract of a  Sinularia, including sesquiterpenes + 394.3316 , [M + Na]+ 416.3128 .Pale yellow oil. [α]24D −9.5° ; UV (PDA) λmax nm: 203, 227; IR νmax cm−1: 3408, 2963, 2926, 1734, 1635, 1455, 1372, 1234, 1024, 904; 1H  and 13C  NMR data m/z [M + Na]+ 369.2396 .Colourless oil. [α]1H-NMR and 13C-NMR spectral data were consistent with published values [m/z [M + Na]+ 345.2398 .Colourless oil. d values ; ESI-FTM1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .1H-NMR and 13C-NMR spectral data were consistent with published values [Colourless oil. d values .R*,5R*,9S*,10R*,11Z)-4-methoxy-9-((dimethylamino)-methyl)-12,15-epoxy-11(13)-en-decahydronaphthalen-16-ol (1) and the lobane -loba-8,10,13(14),15(16)-tetraen-17,18-diol-17-acetate (2), together with the eight known compounds sarcophytol-B [E,3E,7E)-11,12-epoxycembratrien-15-ol [R)-loba-8,10,13(15)-trien-17,18-diol [Sinularia sp. Although there are many publications detailing the isolation of lobanes [Sinularia, this report shows that new investigations are still yielding further new and somewhat unprecedented derivatives, and that continued investigations of this genus are warranted. The biological and pharmacological properties associated with soft coral chemistry, in particular terpenoids, have been shown to be highly promising [Two new compounds, the somewhat unprecedented nitrogen containing (4phytol-B , (1E,3E,en-15-ol , 14,18-een-15-ol , lobatrien-15-ol  and (17R,18-diol , were isromising , leading


Dibenzoazepine (DB) derivatives are important and valuable compounds in medicinal chemistry. The synthesis and chemotherapeutic properties of naturally occurring DBs and different heterocyclic moiety tethered DBs are reported. Herein, we report the DB-fused hybrid structure that containing isoxazolines (DBIs) and their anti-cancer activity, which could throw light on the structural and functional features of new molecules.4g (5--5 H-dibenzoazepine), was found to inhibit the invasion of LM8G7 cells strongly, when compared to other structurally related compounds. Cumulatively, the compound 4g inhibited the invasion MDA-MB-231 cells completely at 10 μM. In addition to anti-invasion property the compound 4g also inhibited the migration of LM8G7 and human ovarian cancer cells (OVSAHO) dose-dependently. Compound 4g inhibited the proliferation of LM8G7, OVSAHO, human breast cancer cells (MCF-7) and human melphalan-resistant multiple myeloma (RPMI8226-LR5) cells that are comparable to cisplatin and suramin.The synthesis and characterization of novel ring DB tethered isoxazoline derivatives (DBIs) were carried out. After the detailed structural characterization using 2D-NMR experiments, the compounds were identified as 5-substituted isoxazolines. The effect of newly synthesized DBIs against the invasion of murine osteosarcoma (LM8G7) cells was studied. Among the tested molecules, compound  N-position play a vital role to augument the biological activity  is one of the synthesized effective anticonvulsant drugs, which is the most frequently prescribed first-line drug for the treatment of epilepsy. D,2. DN-poepilepsy. DB-baseepilepsy,8. Azepiepilepsybenzodiazepilepsy,10. Thesepilepsy,12. Syntepilepsy. SeveralNucleosides are the building blocks of DNA and become a key molecule in the field of medicinal chemistry for the discovery of new natural nucleoside derivatives. So, extensive modifications have been performed on both the heterocyclic base as well as on the sugar moiety. Mainly, the replacement of the ribose ring with an isoxazolidine nucleus has emerged as an interesting class of dideoxynucleoside analogues-16. ThesIn continuation of our effort to synthesise novel DBs and isoxazoli(di)ne derivatives-21 herei2 was prepared by the reaction of commercially available iminostilbene 1 with allylbromideazepine for the first time. After the detailed structural characterization using 2D-NMR experiments, the products were confirmed as 5-substituted isoxazolines. Among the tested compounds, compound 1 H, 13C, HMBC, HMQC were carried out at 500 (125) MHz and the reported chemical shifts (δ) are given in ppm and the coupling constants (J) in Hertz (Hz). Multiplicities of NMR signals are designed as s (singlet), d (doublet), m . Mass spectra were recorded on a Trio 1000 Thermo Quest spectrometer in the electron impact mode or a Platform Micromass spectrometer in the electro spray mode. TLC was performed on silica gel Alugram SilG/UV254 (Macherey-Nagel). The murine osteosarcoma cell line LM8G7, a highly metastatic murine osteosarcoma cell line with the potential to form tumor nodules in the liver, was cloned from LM8G5 cells as described.The compound 1 H NMR : δ 4.4 , 5.12 , 5.28 , 5.8 , 6.76 , 7.08 , 6.98–7.12 , 7.2–7.3 .IR KBr (cm-1): 1315, 1642, 3040, 3072. Anal. Calcd for C17H15N: C, 87.6; H, 6.49; N, 6.0. Found: C, 87.63; H, 6.43; N, 6.0.The product is yellow solid. mp: 40–42°C. 3 (2 eq) was dissolved in 10 mL of methylene dichloride taken in a round bottom flask. The solution was made basic using triethylamine (0.05 eq). 2 equivalents of sodiumhypochlorite, followed by tricyclicamine (1 equiv) 2, were added slowly at 0°C to the stirring solution. After complete addition of the reactants stirring was continued until the reaction is completed. The crude residue was purified by silica gel column chromatography eluting with a mixture of n-hexane and ethyl acetate with successive increasing the quantity of ethyl acetate, affording the corresponding product 4(a-h).The weighed amount of aldoxime 1 H NMR : δ 3.28 ; 3.37 ; 3.51 ; 4.33 ; 4.88 ; 6.76 ; 7.1-8.20 . 13C NMR : δ 38.2 (C-4), 55.6 (C-6), 77.5 (C-5), 124 (CH), 127–144.0 (Ar-C), 158.2 (C-C = N). MS (ESI + ion): m/z = 398.1 [M + H] +. Anal. calcd for C24 H19 N3O3: C, 72.53; H, 4.82; N, 10.57. Found : C, 72.45; H, 4.86; N, 10.48.The product is a thick liquid. Yield: 0.229 g (67.3%).1 H NMR : δ 3.24 ; 3.37 ; 3.45 ; 4.33 ; 4.77 ; 6.70 ; 7.4-8.26 . 13C NMR : δ 38.6 (C-4), 54.3 (C-6), 76.4 (C-5), 125.2 (CH), 130–146.2 (Ar-C), 154.8 (C-C = N). MS (ESI + ion): m/z = 398.6 [M + H] +. Anal. calcd for C24 H19 N3O3: C, 72.53; H, 4.82; N, 10.57. Found : C, 72.48; H, 4.78; N, 10.41.The product is a thick liquid. Yield: 0.224g (65.7 %). 1 H NMR : δ 3.21 ; 3.30 ; 3.48 ; 3.76 ; 3.79; 4.28 ; 4.84 ; 6.78 ; 6.82-7.42 . 13C NMR : δ 37.5 (C-4), 54.7 (C-6), 56.4 (OCH3), 76.9 (C-5), 126 (CH), 128–134.0 (Ar-C), 156.6 (C-C = N). MS (ESI + ion): m/z = 443.5 [M + H] +. Anal. calcd for C27 H26 N2O4: C, 73.2; H, 5.92; N, 6.33. Found : C, 73.15; H, 5.86; N, 6.28.The product is a thick liquid. Yield: 0.260 g (68.6%). 1 H NMR : δ 3.24 ; 3.36 ; 3.50 ; 3.72 ; 4.30 ; 4.88 ; 6.64 ; 6.82-7.32 . 13C NMR : δ 36.2 (C-4), 53.8 (C-6), 56.8 (OCH3), 76.2 (C-5), 126.8 (CH), 128–136.0 (Ar-C), 158.1 (C-C\= N). MS (ESI + ion): m/z = 383.75 [M + H] +. Anal. calcd for C25 H22 N2O2: C, 78.51; H, 5.80; N, 7.32. Found : C, 78.58; H, 5.89; N, 5.68.The product is a thick liquid. Yield: 0.214g (65.3 %). 1 H NMR : δ 2.88 ; 3.28 ; 3.39 ; 4.27 ; 4.72 ; 6.70 ; 6.52-8.22 . 13C NMR: δ 36.4 (C-4), 51.8 (C-6), 75.51 (C-5), 124.22 (CH), 124–152.9 (Ar-C), 158.21 (C-C = N). MS (ESI + ion): m/z =354.0 [M + H] +. Anal. calcd for C23 H19 N3O: C, 78.16; H, 5.42; N, 11.89. Found : C, 78.21; H, 5.34; N, 11.78.The product is thick liquid. Yield: 0.22 g (72.6 %). 1 H NMR : δ 3.11 ; 3.28 ; 3.48 ; 4.28 ; 4.84 ; 6.73 ; 7.00-7.33 . 13C NMR : δ 36.10 (C-4), 51.25 (C-6), 75.81 (C-5), 121 (CH), 122–131.0 (Ar-C), 153.00 (C-C = N). MS (ESI + ion): m/z =353.1 [M + H] +. Anal. calcd for C24 H19 N2FO: C, 77.82; H, 5.17; N, 7.56. Found : C, 77.9; H, 5.21; N, 7.48.The product is yellow thick liquid. Yield: 0.23 g (72.3 %). 1 H NMR : δ 3.15 ; 3.20 ; 3.45 ; 4.30 ; 4.29 ; 6.73 ; 7.00-7.52 . 13C NMR : δ 38.54 (C-4), 53.78 (C-6), 78.71 (C-5), 124 (CH), 127–135.9 (Ar-C), 155.56 (C-C = N). MS (ESI + ion): m/z =387.0 [M + H] +. Anal. calcd for C24 H19 N2ClO: C, 74.51; H, 4.95; N, 7.24. Found : C, 75.08; H, 5.14; N, 7.08.The product is yellow solid. Yield: 0.25 g (75 %). mp-156–158°C. 1 H NMR : δ 3.11 ; 3.24 ; 3.48 ; 4.31 ; 4.71 ; 6.68 ; 6.8-7.62 . 13C NMR : δ 34.70 (C-4), 51.80 (C-6), 76.61 (C-5), 123.75 (CH), 128.8-165.9 (Ar-C), 158.80 (C-C = N). MS (ESI + ion): m/z =389.14 [M + H] +. Anal. calcd for C24 H18 N2F2O: C, 74.21; H, 4.67; N, 7.21. Found : C, 74.11; H, 4.54; N, 7.10.The product is thick liquid. Yield: 0.209 g (69.2 %). in vitro invasion assay was performed using bio-coat Matrigel invasion assay system , according to the manufacturer’s instructions. MDA-MB-231 cells (2 × 105 cells) or LM8G7 cells (2.5 x 104) were suspended in serum-free RPMI 1640 medium or DMEM, respectively and seeded into the Matrigel transwell chambers consisting of polycarbonate membranes with 8-μm pores[The μm pores. After iTM chamber (8 μm PET pores) without Matrigel  in vitro[4) were prepared by detaching and resuspending the cells in DMEM containing 0.1 % BSA. Before the cells were added, the chambers were rehydrated for 2 h in an incubator at 37°C. The lower chambers were filled with DMEM containing 5 % FBS. To the upper chamber, LM8G7 or OVSAHO cells and compound 4g in serum-free DMEM was added. After incubation for 24 h, cells that had passed through the BD BioCoatTM chamber and remained attached to the opposite surface of the membrane are stained with Diff-Quick solution and counted in five random microscopic fields per filter. The % inhibition of the migration of LM8G7 or OVSAHO cells by compound 4g was calculated.The efficacy of LM8G7 cell migration was assessed using the BD BioCoat in vitro. BrieflyTM system . Briefly, LM8G7 cells (5x104 cells) were added to ACEA 96X microtiter plates (e-plateTM) in 100 μl of medium[4g (2 to 75 μM), in 150 μL of DMEM were individually added. The effect of the compounds on the proliferation of LM8G7 or OVSAHO cells was continuously monitored up to 48 hours for every 10 min. The proliferation was monitored for a period of 70 h and expressed as a cell index (quantitative measurement of cell proliferation) as per the manufacturer’s instructions. A cell index was plotted against time (for duplicate experiments). The IC50 values were calculated from concentration-response curves by a non-linear regression analysis using the GraphPad Prism .The cell proliferation assay was done using the RT-CESf medium. In an e4g on MCF-7, melphalan-resistant multiple myeloma (RPMI8226/LR5) MCF-10A or UV♀2 cells was determined by MTT dye uptake method as described previously[4g in a final volume of 0.2 ml for different time intervals at 37°C. Thereafter, 20 μL MTT solution (5 mg/ml in PBS) was added to each well. After a 2 h incubation at 37°C, 0.1 ml lysis buffer  was added; incubation was continued overnight at 37°C; and then the optical density (OD) at 570 nm was measured by Tecan plate reader.The anti-proliferative effect of the compound eviously. Briefly6 MCF-7 cells were pretreated with compound 4g (30 μM) for various time points and then subjected to annexin V staining. Cells were washed, stained with FITC-conjugated anti-annexin V antibody, and then analyzed with a flow cytometer [One of the early indicators of apoptosis is the rapid translocation of the membrane phospholipid phosphatidylserine from the cell’s cytoplasmic interface to the extracellular surface and its accumulation there, producing a loss of membrane symmetry that can be detected using annexin V. Briefly, 1 x 10ces, US).MPS and Basappa lead the medicinal chemistry efforts for the project and assisted in writing the manuscript. Basappa and MS conceived the project, established the in vitro assays, and participated in the experimental design and wrote the paper. SN, AC and DSP are responsible for experimental design of the study. GS was responsible for experimental design of the study, supervised KM, and FL, carried out the invasion assays and proliferation assays . KS and KSR designed the experiments, supervised MPS and Basappa. All authors read and approved the final manuscript.