Src-associated protein of 68 kDa in mitosis (Sam68) is known to bind RNA and be involved in cellular processes triggered in response to environmental stresses, including virus infection. Interestingly, Sam68 is a multi-functional protein implicated in the life cycle of retroviruses and picornaviruses and is also considered a marker of virus-induced stress granules (SGs). Recently, we demonstrated the partial redistribution of Sam68 to the cytoplasm in FMDV infected cells, its interaction with viral protease 3Cpro, and found a significant reduction in viral titers as consequence of Sam68-specific siRNA knockdowns. Despite of that, details of how it benefits FMDV remains to be elucidated.The nuclear protein Sam68 cytoplasmic localization was examined by immunofluorescent microscopy, counterstaining with antibodies against Sam68, a viral capsid protein and markers of SGs. The relevance of RAAA motifs in the IRES was investigated using electromobility shift assays with Sam68 protein and parental and mutant FMDV RNAs. In addition, full genome WT and mutant or G-luc replicon RNAs were tested following transfection in mammalian cells. The impact of Sam68 depletion to virus protein and RNA synthesis was investigated in a cell-free system. Lastly, through co-immunoprecipitation, structural modeling, and subcellular fractionation, viral protein interactions with Sam68 were explored.pol and 3Cpro consistent with early observations of FMDV 3Cpro-induced cleavage of Sam68.FMDV-induced cytoplasmic redistribution of Sam68 resulted in it temporarily co-localizing with SG marker: TIA-1. Mutations that disrupted FMDV IRES RAAA motifs, with putative affinity to Sam68 in domain 3 and 4 cause a reduction on the formation of ribonucleoprotein complexes with this protein and resulted in non-viable progeny viruses and replication-impaired replicons. Furthermore, depletion of Sam68 in cell-free extracts greatly diminished FMDV RNA replication, which was restored by addition of recombinant Sam68. The results here demonstrated that Sam68 specifically co-precipitates with both FMDV 3Dpro and 3Dpol and showed that mutations at RAAA motifs in IRES domains 3 and 4 cause a decrease in Sam68 affinity to these RNA elements and rendered the mutant RNA non-viable. Interestingly, in FMDV infected cells re-localized Sam68 was transiently detected along with SG markers in the cytoplasm. These results support the importance of Sam68 as a host factor co-opted by FMDV during infection and demonstrate that Sam68 interact with both, FMDV RNA motifs in the IRES and viral non-structural proteins 3Cpro and 3Dpol.We have found that Sam68 is a specific binding partner for FMDV non-structural proteins 3CThe online version of this article (doi:10.1186/s12985-015-0452-8) contains supplementary material, which is available to authorized users. Foot-and-mouth disease virus (FMDV) is one of the most contagious animal viruses and causes disease in important livestock species including cattle, pigs, and sheep. The virus is represented by seven different serotypes: A, O, C, Asia1, SAT1, SAT2, and SAT3; each with a distinctive global distribution. Control of FMD outbreaks are typically combated through a combined strategy of mass vaccination and animal culling. Given the profound economic fallout following the spread of FMDV, it is of critical importance to understand the factors that influence FMDV virulence in order to design improved anti-viral strategies [Picornaviridae family. It possesses a single-stranded positive-sense RNA genome encoding a single polyprotein that is subsequently cleaved to produced four structural proteins  and ten non-structural proteins . The genomic region coding for polyprotein is flanked by 5′ and 3′ non-translated regions (NTRs) [trans-acting factors (ITAFs) also impact IRES activity. FMDV contains a type II IRES that is predicted to fold into five domains, followed by a polypyrimidine tract and by two alternative AUG initiation codons [FMDV is the prototypic member of the Aphthovirus genus of the s (NTRs) , functios (NTRs) –10. IRESn codons . GNRA mon codons .pro). Interestingly, Sam68 knockdown resulted in a more significant reduction of FMDV titers than its impact in IRES-driven protein translation when using a bicistronic construct [In a previous study, we presented evidence suggesting that the 68 kDa Src-associated protein in mitosis (Sam68) host protein binds to the IRES elements of FMDV, which could impact IRES translation activity. We had also shown appearance of Sam68 cleavage products in FMDV-infected cells and in the presence of semi-purified recombinant 3C proteinase : T-cell intracellular antigen 1 (TIA-1), in agreement with reports for Sam68 being a stress granule  marker fIn our previous study, FMDV infection was shown to induce a partial redistribution of the nuclear protein Sam68 to the cell cytoplasm in the LFBK cell line , 55, 56.Next we sought to determine if cytoplasmic Sam68 localized with specific SG markers as reported for poliovirus (PV) . To test24 IRES (predicted using M-fold software). Figure The initiation of cap-independent translation by IRES elements in the 5′ NTR of the picornavirus genome is regulated by RNA-RNA and RNA-protein interactions with canonical and non-canonical translation initiation factors , 62–66. 24-Cru IRES nt 435–499) encoding the wild-type (WT) sequence or double mutated forms (UAAA to UACG), as described in Material and Methods and Table 50) in the range of 2–5 μM, under the experimental conditions. Interestingly, although the mutant domain 4 probe also produced RNP in the presence of Sam68, this binding required approximately 10-fold higher amounts of the protein to produce the same amount of RNP products as with the WT domain 4 probe  long RNA probes  was expressed and tested side-by-side with full-length protein in an EMSA assay. To this end, EMSA experiments were carried out using WT domain 4 probe (as described above) and either an intact (Sam68-WT) or mutant Sam68 (Sam68-KH-del),  showed no differences between reactions programed with non-depleted, depleted (only) and depleted CFE supplemented with 1 μM Sam68-6H  FMDV replicons. The full genome WT or I3 (CAAA to CACG), I4 (two UAAA to UACG), or the two combined (I34) mutant plasmids were first linearized by SwaI restriction enzyme digestion and then transcribed into RNA, and electroporated into BHK-21 cells. Twenty-four hours later, the transfected cells were freeze-thawed, and a fraction of the lysate (1/10th) was used to initiate next passage. In contrast to the WT virus that exhibited visible cytopathic effect in the second passage in BHK-21 cells, no viable virus was recovered from any of the mutant RNAs even after 7 blind passages (data not shown). This was a surprising observation that led to suspicions about the ability of the mutant RNAs to translate and/or replicate. The same panel of IRES mutations were inserted into FMDV A24 G-luc replicons  or mutant  sequences, were digested with SwaI restriction enzyme prior to the RNA transcription. The RNA were then transfected into BHK-21 cells and samples were collected at 0, 2, 6 and 24 h post transfection (hpt) and examined for G-luc expression. As shown in Fig. 24G-luc plasmids using MfeI restriction digests . The results in Fig. To understand the functional significance of Sam68-binding to the FMDV IRES, we next examined the effect of a CAAA to CACG mutation in domain 3 and on two UAAA to UACG mutations in domain 4 using FMDV Aons Fig.  to evaluons Fig. . Plasmidpol as had previously been shown for poliovirus [pol peptides binders, and (iii) structural prediction and molecular docking of Sam68 and FMDV 3Dpol  crosslinked to protein G Dynabeads. As a control, lysates of virus-infected cells were subject to IP with an isotype antibody control crosslinked to protein G as described above. The left panel of Fig. pol IP developed with anti-Sam68. In lane 1, a control IP with isotypic antibody shows the absence of a Sam68 specific band. The two faint bands of approximately 20 and 30 kDa are non-specific proteins pulled down by this antibody. Lanes 2 and 3 are the control cell lysates at 1:10 dilution from mock-infected and FMDV-infected cell lysates, respectively. As expected, the endogenous Sam68 as well as the previously described 55 kDa Sam68-like-mammalian protein 2 (SLM-2) [pol IP pulled down Sam68 specific protein band of 68 kDa and three additional Sam68 reactive bands. In addition to the proteins of expected size in the IP experiments, higher molecular weight Sam68 reactive bands in FMDV 3Dpol IP could be attributed to a shift due post-translational modifications of Sam68 [pol and other cellular proteins to Sam68 thus producing the higher molecular weight protein bands. The right panel of Fig. pol. The isotype antibody IP (lane 1) and mock-infected cell lysates (lane 2) showed no 3Dpol specific band. In contrast, lane 3 (FMDV-infected cell lysates control) and lane 4 (anti-Sam68 IP) showed 3Dpol specific bands. Together, the reciprocal pulldown experiments of Sam68 and FMDV 3Dpol from FMDV-infected cell lysates provides the evidence for Sam68 and FMDV 3Dpol interaction in virus-infected cells.For the co-immunoprecipitation analysis, BHK-21 cells were either mock infected or infected with FMDV at a MOI of 10 Fig. . Five ho (SLM-2) , 73 wereof Sam68 . The lowof Sam68 . Alternaof Sam68 , 74 may pol that interacts with Sam68, we probed for binding of Sam68 in an ELISA using a 3D peptide fragment library consisting of fragments of about 50 amino acids (aa) in length starting from the N-terminus of 3Dpol . As evident from the data, frag-4 and frag-8 of 3Dpol bind to Sam68 with high affinity whereas frag 2, 6 and 7 bind to Sam68 with low affinity -Sam68 (blue) complex in cartoon mode. Frag-4 residues 193–217 shown in orange color and frag-8 residues 453–470 in red color form the major Sam68 binding interface in 3Dpol. Although, frag-5 residues 221, 222, 225, 226 also align the Sam68 binding interface, the major participation in the binding comes from frag-4 and 8, consistent with the peptide library ELISA data, further reinforcing the importance of these two regions in FMDV 3Dpol in interaction with Sam68. The fact that Sam68 has been shown to interact with other picornaviruses and that 3Dpol structure is largely preserved among picornaviruses, there might exist a common mechanism by which Sam68 affects picornavirus replication. Together the data from IP, ELISA and computational docking suggest that Sam68 binds to FMDV 3Dpol including residues contained in frag-4 and frag-8.Furthermore, we used molecular docking software to illustrate the putative interaction between 3Dm68 Fig. . Panels pro as infection progressed, we wanted to ascertain if Sam68 could be similarly co-precipitated with 3Cpro as was observed with 3Dpol. Mock-infected or FMDV-infected cells at a MOI of 10 were lysed at 1, 3, and 5 hpi. The resulting cell lysates were subject to IP with mouse monoclonal anti-FMDV 3Cpro, and the eluates thereof were probed with rabbit polyclonal anti-Sam68  on a Western blot. A strongly reacting band at 68 kDa was detected at 5 hpi with a few lower molecular weight fragments faintly reacting, as well as a faint 68 kDa band at 3 hpi , which revealed the appearance of Sam68 cleavage fragments at 5 hpi coincident with the binding interaction between Sam68 and 3Cpro  proteins and to evade the host defense mechanisms . The assin vitro translation-RNA replication assays, which have proven to be instrumental in defining the role of many individual host or virus proteins at the molecular level [in vitro cell-free experiment using Sam68-depleted extracts, revealed a defect in FMDV RNA synthesis, but not a significant reduction on virus translation, which we did not anticipate based on earlier studies [pol by Western blot. Future studies using lower RNA concentration and shorter incubation times to program FMDV protein synthesis in CFEs could help resolve this issue.In light of the above-mentioned observations, we sought to explore the functional significance of the Sam68-FMDV IRES interaction in more detail. To that end, we first utilized cell-free ar level –91. Our  studies , 92. We pro and 3Dpol in infected cells. PV 3Dpol has also been shown to interact with Sam68 [pol fragments, it was suggested that 3Dpol frag-4 (aa 158–217) and frag-8 (aa 405–470) bind Sam68 with high affinity. However, under the experimental conditions, we cannot exclude the possibility that frag-2 (aa 49–108), frag-6 (aa 269–331), and frag-7 (aa 332–404) could also provide a Sam68 binding interface. In fact, the docking poses of the electrostatic surfaces of FMDV 3Dpol [Fig. pol is formed by aa 193–217 and aa 453–470 in frag-4 and 8 that are part of the functionally critical palm and thumb domains of 3Dpol Fig. pol structure consisting of thumb, palm, finger and fingertips domains is conserved among picornaviruses. Another striking feature of the Sam68-3Dpol interaction is the charge complementarities between the binding surfaces of the two proteins (electro-negative of 3Dpol and electro-positive of Sam68). Further studies will be required to determine the significance of these protein interactions for viral infection.Results obtained in our characterization of mutated forms of genetically engineered G-luc replicons, as well as viral genomes, provide compelling evidence for the significance of the RAAA motifs in domain 3 and 4 in FMDV IRES-driven translation. It is noteworthy that the full genome and replicon mutants exhibited impairment in their translation and replication. Interestingly, our earlier studies showing reduction in FMDV titers by Sam68 siRNA knockdown and the results described herein, suggest that it is possible that subtle modifications in the Sam68 interaction with the FMDV IRES could impact other functions that this protein exerts that are needed for efficient virus replication. This supposition is in line with multi-functional properties attributed to Sam68 see . Indeed,th Sam68 . Using apro and 3Dpol also raises additional questions regarding the FMDV-induced cleavage of Sam68. The FMDV 3Dpol and transiently expressed 3CD precursor are known to partially localize to the cell nucleus due to a nuclear localization signal in the N-terminus of 3Dpol [pro from nuclear-localized 3CD precursor [pro cleavage of host cell transcription factors found in the nucleus of PV-infected cells also supports this notion [pro and 3Dpol, it remains to be determined whether WT full-length Sam68 is cleaved by FMDV 3CD or fully matured 3Cpro. Moreover, since we also observe by Western blot some accumulation of full-length Sam68 in the cytoplasm as FMDV infection progresses, it is undetermined whether the full-length or cleaved form of Sam68 contributes to the modulation in virus replication. Like the full-length Sam68, the 3Cpro cleaved Sam68 is predicted to maintain its RNA-binding KH domain. Potentially, the C-terminal cleavage eliminates steric hindrances allowing for tighter binding.The observation that Sam68 co-precipitates with both the FMDV 3Cof 3Dpol –95. Thisrecursor . The 3Cps notion . Therefoin vitro in the cell-free system. Interestingly, Piotrowska et al. [The significance of the appearance of Sam68 truncation products as FMDV infection progresses still remains to be determined, yet the significance of Sam68 to HIV infection has been highlighted by how a Sam68 isoform with a C-terminal deletion (Sam68DeltaC) potently inhibits the progression of HIV infection , 30. Sama et al.  postulata et al. . Given tin vitro. Disruption of the IRES RAAA motif leads to non-viable virus progeny and defective replicons, though this may not be attributable to Sam68 interaction with the IRES alone. Finally, consistent with a role in RNA replication and its observed cleavage during FMDV infection, Sam68 co-precipitates with both FMDV 3Cpro and 3Dpol.Sam68 is a host factor co-opted by FMDV during infection of susceptible host cells. As infection progresses, Sam68 becomes partially redistributed to the cytoplasm, where earlier in infection, it co-localizes with markers of SGs. Interaction of Sam68 with the FMDV IRES appears to promote viral RNA translation Stratagene QuickChange IIXL mutagenesis kit was purchased from Agilent Technologies, Inc. . Lipofectamine 2000 DNA transfection reagent and streptavidin-coupled magnetic beads were purchased from Invitrogen . Sam68 expression plasmids pGEX-2 T Sam68 (Containing GST-tagged Sam68), pcDNA3 HA-tagged Sam68-WT and pcDNA3 HA-tagged Sam68 delta-KH (Sam68-KH-del) were purchased from Addgene Cambridge, MA, USA. LightShift® Chemiluminescent RNA EMSA Kit and the NE-PER cellular fractionation kit were purchased from Thermo Fisher Scientific Inc. . Restriction enzymes EcoRI and BamHI were purchased from New England Biolabs .pol antibody was a gift from Dr. A. Clavijo at the National Centre for Foreign Animal Disease, Winnipeg, Manitoba, Canada. Rabbit polyclonal anti-VP1 sera were courteously provided by Dr. M. Grubman, ARS, USDA. Goat-anti-rabbit antibodies conjugated to AlexaFluor-488 (AF-488), donkey-anti-goat antibodies conjugated to AF-568 and goat-anti-mouse antibodies conjugated to AF-568 were purchased from Molecular Probes .Polyclonal rabbit anti-Sam68 directed against the C-terminus of Sam68 (C-20), polyclonal goat anti-TIA-1, and HA-probe Antibody (F-7) were purchased from Santa Cruz Biotechnology . Mouse monoclonal anti-G3BP was purchased from Abcam . Polyclonal rabbit anti-Sam68 directed against the N-terminus of Sam68 was purchased from Novus Biologicals . Monoclonal mouse anti-FMDV 3D24 Cruzeiro/Brazil 1955  was derived from the infectious cDNA clones pA24Cru [24 Cru carrying modified IRES  in Dulbecco’s minimal essential media (DMEM) supplemented with antibiotic/antimycotic [2 atmosphere. All of the PCR primers used to generate different clones and mutations were purchased from Life Technologies . The primer sequences have been detailed in Table E. coli expression.FMDV type A pA24Cru . FMDV A2imycotic . Cells wMammalian cells were transfected with plasmid DNA using the Lipofectamine 2000 transfection reagent (Invitrogen) following the manufacturer’s protocol. Briefly, cells were grown to approximately 30–40 % confluence. The cell culture medium was replaced with serum-free media containing the desired plasmid and Lipofectamine 2000. Cells were incubated with the mixture for 48 h at 37 ° C. Post-incubation, the cells were harvested for examination by Western blot to confirm expression of the desired construct.24 Cru at a MOI of 10 and incubated at 37 °C for 1 h. Excess virus was removed by acid-washing the cells briefly followed by three rinses with virus growth media (VGM). Designated cells were then fixed with 4 % PF, representing a 1 hpi time point. Remaining cells were provided fresh VGM and incubated for 2, 3, 4, or 5 hpi at 37 °C, after which, they were fixed with 4 % PF. Cells were washed in PBS, permeabilized with 0.1 % Triton-X100 in PBS for 20 min on ice, and blocked with 3 % bovine serum albumin (BSA) for 30 min at RT. The cells were probed with primary antibodies at RT for 1 h. The following day the cells were incubated for 1 h at RT with goat-anti-rabbit and goat-anti-mouse conjugated with AF-488 or AF-568 (Molecular Probes), respectively. The cover slips were washed three times with PBS after each antibody treatment. After the final antibody treatment, the cover slips were air-dried and mounted onto glass slides with ProLong antifade medium supplemented with DAPI stain (Life Technologies). Cells were examined and images captured using a 100X oil immersion objective on an Olympus fluorescent microscope . Images were refined and figures generated using Adobe Photoshop software . When examining cells transfected with GFP-tagged Sam68, the permeabilization and subsequent antibody steps were omitted. Following fixation and PBS washes, the coverslips of the transfected cells were mounted using ProLong anti-fade with DAPI and examined as described above.Cells were grown on glass coverslips in 12-well plates until approximately 40 % confluence. Control uninfected cells were fixed immediately with 4 % paraformaldehyde  in PBS fSDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) was carried out using a 12 % Nu-PAGE® pre-cast gel system (Invitrogen). Subsequently, the separated proteins were electro-blotted onto a nitrocellulose membrane . After blocking the membrane with 5 % milk in PBS-T, specific proteins were detected with primary antibodies at indicated dilutions followed by goat-anti-rabbit or goat-anti-mouse antibodies conjugated with HRP (Sigma-Aldrich). Cellular α-tubulin, employed as an internal loading control protein, was detected with mouse monoclonal anti-tubulin antibody  that in turn was detected with  goat anti mouse-HRP (Sigma-Aldrich). The bound HRP conjugate antibodies were reacted with the WestDura SuperSignal chemiluminescent reagent (Pierce) according to the manufacturer’s instructions and visualized on X-ray film .In order to generate hexahistidine (6H) fused-Sam68 (Sam68-6H), the Sam68 coding gene was sub-cloned into pET30c + plasmid from pGEX-2 T Sam68 plasmid using BamHI and EcoRI restriction sites. Rosetta-DE3 cells were transformed with Sam68-6H-pET30c + plasmid. Single colony containing the plasmid was inoculated in 100 mL LB broth containing 0.4 % glucose, 25 μg/mL kanamycin (K25) and 20 μg/mL chloramphenicol (C20), and grown at 30 ° C with 220 rpm rotor speed in a Thermo Scientific MaxQ 4000 benchtop orbital shaker. This overnight culture was used as a starter culture for 1 l (L) LB K-25, C-20 broth at 37 ° C. At an OD of 0.8, the culture was induced with 1 mM IPTG for 3 h. The cell pellets were harvested by centrifugation at 4000 rpm at 4 ° C in a Sorvall high-speed centrifuge. Sam68-6H was lysed for 20 min at RT with intermittent stirring in BugBuster™ cell lysis reagent supplemented with pepstatin and leupeptin. To disrupt the cellular nucleic acids cell lysate was then sonicated at 85 % slope for 5 min at 30 s intervals. After centrifugation of the lysates for 30 min at 12,500 rpm at 4 ° C, the supernatant was mixed with 5 mL Ni-NTA agarose slurry pre-equilibrated with 6H-binding buffer (Pierce) and incubated at RT with gentle shaking. This slurry was packed in a polypropylene column and eluted with 5 column volumes of 6H-elution buffers (Pierce). The fractions with highest protein purity were pooled and dialysed against 50 mM Tris–HCl pH 8.0 containing 10 mM β-mercaptoethanol, 10 % glycerol, pepstatin and leupeptin. Protein concentration was monitored against reagent blank at 280 nM and the identity of protein was confirmed by Western blot using 1:500 rabbit polyclonal antibody raised against C-terminus of Sam68 .HEK-293 cells containing SV40 T-antigen (293 T cells) were either transfected with pcDNA3 HA-tagged Sam68-WT or pcDNA3 HA-tagged Sam68 delta-KH (Sam68-KH-del) using Lipofectamine-2000 reagent. After 48 h, the cells were lysed by incubating for 5 min in 1X NP40 lysis buffer containing 50 mM Tris–HCl, pH 8, 150 mM NaCl and 1 % NP40. Following lysis, the debris were removed by centrifuging the lysate at 15,000 rpm at 4 ° C and 20 min. HA-tagged Sam68-WT or Sam68-KH-del were purified using HA-tagged protein purification kit  as per manufacturer’s instructions. The purified proteins were diluted to 10 times their initial volume in 1X RNA-EMSA buffer  and concentrated using Amicon Ultra-15 Centrifugal Filter Units with 30 KDa cutoff . The identity of proteins was confirmed by Western blot probing with HA-probe antibody . Purified HA-tagged Sam68-WT and Sam68-KH-del were stored at −80 ° C until further use.24 Cru infectious clone linearized with restriction enzyme SwaI was utilized as template for generating various IRES domain RNAs. Full-length IRES was amplified with primer pair P1579 S IRES -T7 and P1580 AS IRES. Various IRES fragments were amplified with their respective primer pairs as following: P1329 A24-T7cre and P1330 AS cre (cre-fragment); sense (S) 5′ NTR T7-P1287 and antisense (AS) 5′ NTR-P1288 (S-fragment); P21036 IRES loop 1-T7 and P21037 IRES loop 1 Reverse (IRES loop 1); P1584 IRES loop 2 Forward-T7 and P1585 IRES loop 2 Reverse (IRES loop 2); P21034 IRES loop 3-T7 and P21035 IRES loop 3 Reverse (IRES loop 3); P-1581-IRES-4 Forward-T7 and P-1582-IRES-4 Reverse (IRES loop 4). See Table FMDV ABiotinylated FMDV IRES full length, domain 2, domains 2–4, and domain 4 RNA were transcribed using the Megascript T7 kit (Ambion) with rNTPs supplemented with biotin-11-CTP (Roche). First, 50 μL of streptavidin-M280 magnetic beads (Invitrogen) were pulled-down by a magnet and equilibrated in 1 mL of 1X binding and washing (B&W) buffer consisting of 5 mM Tris–HCl, pH 7.5, 0.5 mM EDTA, and 1 mM NaCl. Following another pulldown and removal of B&W buffer, 5 μg of recombinant Sam68 and 100 μg of tRNA were added to the beads and incubated at 37 ° C for 30 min in a final volume of 200 μL 1X B&W buffer. The beads with potentially bound RNA and protein were pulled-down and washed three times in B&W buffer. Finally, the beads were resuspended in 20 μL of 1X Laemmli sample buffer, heated at 98 ° C for 10 min, and the supernatant collected after centrifugation was loaded onto a 12 % SDS-PAGE gel followed by Western blot analysis probing with anti-Sam68 (Santa Cruz Biotechnology).2, 10 % glycerol, 1.0 mM DTT, 1.0 μg/μL tRNA and different concentrations of recombinant Sam68. Protein bound RNA was resolved from the free RNA on 6 % acrylamide gel in 0.5 X TBE buffers for 90 min at 60 volt (V) and 4 ° C, which was pre-run for 1 h at 60 V to maintain uniform current. RNA were transferred from gel to precharged Hybond nylon membrane at 30 V (constant voltage) for 45 min using NuPAGE® Novex®-trans-blot apparatus (Life Technologies) in the presence of 0.5 X TBE buffer. After the RNA were transferred to the nylon membrane, the RNA were crosslinked to the membrane using 2000 MJ-cm−2 ultraviolet radiation twice in Stratalinker UV-crosslinking system (Stratagene). The RNA was detected with LightShift Chemiluminescent EMSA Kit (Pierce) following the manufacturer’s instructions.Sixty five nucleotide long 5′-biotin labeled synthetic RNA probe containing two potential Sam68 binding (UAAA) motifs in WT form (labeled as WT domain 4 probe) or mutated form (UACG) labeled as (mutant domain 4 probe) were purchased from Integrated DNA Technology  (See Table pro or 3Dpol or Sam68 were separately bound and crosslinked to the Protein G Dynabeads (Novagen) using the reagents provided in the Pierce Crosslink Co-immunoprecipitation Kit (Pierce). BHK-21 cells uninfected or infected with FMDV for 4 h were lysed with 1X RIPA buffer  supplemented with proteinase inhibitors and Benzonase™ . Approximately 1–5 mg of uninfected or virus-infected cell lysates were then mixed with different sets of antibody-coupled beads, washed, and eluted. IP eluates were mixed with Laemmli sample buffer [pol.Following the manufacturer’s protocol, antibodies directed against FMDV 3Ce buffer  boiled, pol fragments were diluted to 10 ng/mL, whereas 6His-tagged full-length 3Dpol was diluted to 500 ng/mL in blocking buffer and added to the plate, followed by incubation at 37 ° C for 1 h on a rotary shaker followed by washing on plate washer (SkanWasher 300 Version B) with 0.01 M PBS and 0.05 % Tween 20 (PBS-T). One hundred μL of anti-6His was added at 1/500 dilution in blocking buffer followed by incubating the plate at 37 ° C for 1 h while shaking. The plate was washed using the plate washer (SkanWasher 300 Version B). One hundred μL of HRP (SureBlue-KPL) was added, and after about 10 min when the blue color developed in the positive wells, the reaction was stopped by adding 100 μL of SureBlue-KPL stop solution and read in the plate reader, Biotek ELx808 at 640 nm.A sandwich ELISA was performed where recombinant Sam68 was diluted in buffer carbonate-bicarbonate (pH 9.6) to obtain 0.05 μg/mL, and 100 μL/well was used to coat Nunc Maxisorp plates (Fisher Scientific). After overnight incubation at 4 ° C, the plates were washed on plate washer (SkanWasher 300 Version B) with 0.01 M PBS and 0.05 % Tween 20 (PBS-T). Next, individual 6His-tagged 3D24 3Dpol using SWISS-MODEL [24 3Dpol and Sam68 were docked with Clus-Pro docking interface [For this purpose, first, the structural model of full-length Sam68 was made using Phyre-2.0 web server  and FMDVSS-MODEL . These interface  and 20 b24 RNA was transcribed from FMDV A24 plasmid linearized with the restriction enzyme SwaI. The CFE of BHK-21 cells were prepared with modifications to the protocol described earlier [2 and 25 °C for 30 min. After this, the nuclease was inactivated with 3 mM EGTA, pH 8. Sam68 depletion was performed by mixing 100 μL of rabbit polyclonal anti Sam68 antibody  with 1 mL of BHK-21 CFE and 50 μL Dynabeads protein G for 2 h at room temperature under gentle rotation. Following incubation, the mix was put on magnetic plate to separate the bead bound material (anti-Sam68 antibody-Sam68 protein). The clear supernatant was labeled as Sam68 depleted CFE, and used later for protein and RNA synthesis experiments. The FMDV A24 RNA synthesis, the effect of Sam68 on this process, and the detection of newly synthesized RNA were carried with following modifications to the protocol described by Pogany and Nagy [32 labeled nucleotides, biotin-UTP was used to detect the newly synthesized RNA, 2) purified viral polymerase was used to initiate the polymerase reaction, and 3) the reaction was carried out at 37 ° C for 5 h. The newly synthesized RNA was spotted on pre-charged Hybond nylon membrane followed by detection of the signal with the Pierce nucleic detection kit following the manufacturer’s protocol. The translation reactions were carried out in CFE containing 20 amino-acyl tRNAs creatinine phosphate, creatine kinase, dTT, rATP, rGTP, rCTP, and rUTP at 32 ° C with aliquots collected at 30 min, 60 min, 90 min and 120 min, which were resolved on 4-12 % SDS gel, and detected by Western blot probing with anti-FMDV 3Dpol.FMDV A earlier , 103. Brand Nagy : 1) inst24Cru full genome plasmid clone was digested with restriction enzymes SanDI and XbaI and the fragments purified. Using Infusion primer pairs P-2182-Infusion-FMDV A24 IRES Forward and P-2183-Infusion-FMDV A24 IRES Reverse . The parental AstEII-MfeI digested DNA fragments containing IRES mutations into a pA24 G-luc replicon system . Briefly, these replicons were derived by replacing the Lpro, and capsid coding sequences with a G-luciferase reporter gene, and later sub-cloning the IRES mutations into the A24 G-luc plasmid. The new constructs were labeled as: I3 G-luc, I4 G-luc and I34 G-luc. The RNAs were derived from either SwaI or MfeI digested plasmids and then transfected into BHK-21 cells using Lipofectamine 2000 transfection reagent (Life Technologies). The cell lysate obtained from samples collected at 0, 2, 6 and 24 hpt were clarified and the G-luc signal was assessed using a Microplate Luminometer. The G-luc signals were normalized to the amount total protein measured in each individual lysate. Two independent transfection experiments were conducted for each of the RNA and the G-luc values are expressed as relative luciferase units (RLU) per nanogram of protein.The IRES mutant FMDV replicons were created by substituting the B


Health protocols need to be guided by current data on survival and benefits of interventions within the local context. Periodic clinical audits are required to inform and update health care protocols. This study aimed to review morbidity and mortality in very low birth weight (VLBW) infants in 2013 compared with similar data from 2006/2007.We performed a retrospective review of patients’ records from a neonatal computer database for 562 VLBW infants. These neonates weighed between 500 and 1500 g at birth, and were admitted within 48 hours after birth between 01 January 2013 and 31 December 2013. Patients’ characteristics, complications of prematurity, and therapeutic interventions were compared with 2006/2007 data. Univariate analysis and multiple logistic regression were performed to establish significant associations of various factors with survival to discharge for 2013.Survival in 2013 was similar to that in 2006/2007 . However, survival in neonates who weighed 750–900 g significantly improved from 20.4% in 2006/2007 to 52.4% in 2013 (p = 0.001). The use of nasal continuous positive airway pressure (NCPAP) increased from 20.3% to 62.9% and surfactant use increased from 19.2% to 65.5% between the two time periods (both p < 0.001). Antenatal care attendance improved from 54.4% to 70.6% (p = 0.001) and late onset sepsis (>72 hours after birth) increased from 12.5% to 19% (p = 0.006) between the two time periods. Other variables remained unchanged between 2006/2007 and 2013. The main determinants of survival to discharge in 2013 were birth weight , NCPAP , necrotising enterocolitis , and mode of delivery, including normal vaginal delivery  and vaginal breech .There was a marked improvement in the survival of neonates weighing between 750 and 900 g at birth, most likely due to provision of surfactant and NCPAP. Provision of NCPAP, prevention of necrotising enterocolitis, and control of infection need to be prioritised in VLBW infants to improve their outcome. In 2013, neonatal mortality accounted for almost 45% of deaths in children younger than 5 years of age  g) than in survivors . Similarly, gestational age was significantly lower in non-survivors (27.4 [2.5] weeks) than in survivors . Temperature at admission was significantly lower in non-survivors than in survivors . The neonate’s sex was not associated with outcome. Other significant predictors of survival at discharge are shown in Tables Results of multiple regression analysis for the whole group are shown in Table The number of VLBW infants is increasing, with an increase of 18.5% of VLBW admissions to the CMJAH in 2013 compared with 2006/2007. Care of these infants should become a health priority in the coming decade. Regular review of outcomes and adjustment of health protocols are essential to ensure the best possible outcome of patients. There was a marginal improvement in the overall survival of VLBW infants in this unit, from 70.2% in 2006/2007 to 73.4% in 2013, but this did not reach statistical significance. These survival rates are similar to those in Korea (6), but remain below those of high-income countries . Different interventions are provided for different birth weights in the CMJAH neonatal unit. In the current study, stratifying for birth weight showed that survival of VLBW infants weighing 750–900 g significantly improved from 20.4% in 2006/2007 to 52.4% in 2013, whereas that for infants <750 g and those who weighed 900–1500 g did not change. The reason for this finding can be attributed to the introduction of NCPAP and INSURE for the group of neonates weighing 750–900 g, whereas the management of neonates in the other weight groups remained unchanged between the two time periods. During 2006/2007, only 6.8% of neonates with HMD who weighed 750–900 g at birth were offered NCPAP compared with 94.8% in 2013. The number of neonates who weighed >900 g with HMD requiring IPPV significantly decreased from 35.1% in 2006/2007 to 24.6% in 2013. Importantly, NCPAP is a cheaper therapeutic option than mechanical ventilation with its attendant difficulties and complications. Based on this markedly improved survival of ELBW infants, perhaps physicians should review the birth weight cut-off for NCPAP and INSURE, and offer this to all neonates with RDS, regardless of their birth weight. Provision of available therapeutic interventions to every neonate, irrespective of birth weight, is a major factor in better survival rates of VLBW infants in high-income compared with low- and middle-income countries [The most significant predictor of survival in the present study in 2013 was birth weight, which is consistent with previous findings in 2006/2007. Other significant predictors of survival in the present study were NCPAP, NEC, resuscitation at birth, maternal HIV status, and mode of delivery. These findings are slightly different to those in 2006/2007 where hypotension, sex, and birth outside a health facility were associated with outcome, but these factors were not associated in the 2013 study. However, NCPAP and NEC remained significantly associated with outcome in both time periods. Other significant differences between the two time periods were an increase in attendance at antenatal care, HMD, and late onset sepsis, with a decrease in neonates who were SGA. The significant increase in the number of neonates with HMD between the two time periods was unexpected, but may be associated with the decrease in the number of neonates who were SGA . DespiteSome authors have suggested that there is little additional benefit in using other predictors of outcome over birth weight in resource-constrained settings . HoweverIn the current study, notably, several factors did not improve over time. Administration of antenatal steroids remained low. Many mothers present late in labour and do not attend antenatal care. Therefore, there may be limited opportunities for obstetricians to administer steroids . MaternaCertain complications of prematurity in the current study were low, including pneumothorax, cystic periventricular leukomalacia, severe peri-intraventricular haemorrhage P-IVH, and ROP. This may reflect the relatively poor survival of ELBW infants, but many of the neonates in the present report were not screened. Therefore, these figures may be under-reported. Reliable information on long-term outcome  is essential. Therefore, protocols must be put in place to ensure that all neonates are screened for ROP and have cranial ultrasound performed. The presence of ROP, IVH, and periventricular leukomalacia can be used as proxy indicators of a long-term poor outcome.The mortality rate of VLBW infants did not significantly change between 2006/2007 and 2013. The greatest improvement in survival was observed in VLBW infants who weighed 750–900 g at birth. This most likely reflects the provision of surfactants and NCPAP to this weight category of infants. Health protocols, including simple, inexpensive interventions , should improve survival to discharge in VLBW infants. Although LOS is not associated with mortality, there is an alarming increase in LOS, and infection control measures must also be prioritised. These interventions will not only improve the survival of VLBW infants but also result in lower costs in the care of these neonates.


We evaluated our initial experience of performing sclerotherapy for symptomatic hepatic cysts using polidocanol foam instead of a liquid sclerosant.Three consecutively registered patients with symptomatic hepatic cysts (one with polycystic liver disease) underwent polidocanol foam sclerotherapy. A pigtail catheter was inserted into the targeted cyst following percutaneous cyst puncture under ultrasound guidance, and the cyst fluid was aspirated. To confirm the absence of communications between the cyst and surrounding hepatic vessels, 3 % polidocanol foam sclerosant was injected at a 1:4 ratio of polidocanol to air following digital subtraction cystography with carbon dioxide. C-arm computed tomography (CT) guidance also was used to monitor foam filling of the targeted cyst. The maximum dose of sclerosant injected per treatment session did not exceed 10 ml. The catheter was kept unclamped overnight for open drainage, and additional sclerotherapy sessions were performed on subsequent days, if needed.Efficient sclerotherapy was achieved with an average of two sessions. The initial mean cyst volume was 1,052 ml, and gradual resolution was observed without recurrence. The mean reduction rate was 97.9 % (97.7–98.3 %), and all cyst-associated symptoms disappeared. The median follow-up period was 17 (range 6–21) months. Although one patient experienced moderate-grade fever and another moderate pain at the puncture site, no major complications were observed.Polidocanol foam sclerotherapy is a safe and effective treatment for symptomatic hepatic cysts. Liquid sclerosants have been studied as alternatives to ethanol during sclerotherapy for symptomatic hepatic cysts –3, becauThe present study was designed to evaluate retrospectively the safety and efficacy of polidocanol foam sclerotherapy for symptomatic hepatic cysts.This study was approved by the institutional review board of our hospital and was conducted after obtaining informed consent from all patients. Three consecutive patients with symptomatic hepatic cysts who were referred for interventional radiology between March 2010 and March 2012 were included. Hepatic cysts were detected and properties of the cyst walls and contents evaluated by US, contrast-enhanced computed tomography (CT)  was then introduced into the cyst through a 9-F peel-away introducer sheath  over a 0.038-in. guidewire, and the cyst contents were aspirated. The aspirated fluid volume was measured, and the specimens were examined bacteriologically and cytologically.Percutaneous cyst puncture through normal hepatic parenchyma was performed using a 19-gauge, 20-cm needle  under US guidance after local anesthesia administration. Contact was confirmed by cystic fluid aspiration and by digital subtraction cystography with 10 ml of iodized contrast medium. An 8.5-F pigtail catheter (Ultrathane2)  with 8 ml of room air [The maximum dose of 3 % polidocanol per treatment session was 10 ml . Sclerotherapy was repeated on subsequent days, if needed, on the basis of daily drainage from the catheter and the initial cyst volume. The catheter was removed 24 h after the final sclerotherapy session.The total sclerosant dose used for each session and the presence and severity of any complications were documented. The targeted cyst volume was calculated by CT before sclerotherapy and routinely performed 1 or 6 month(s) after sclerotherapy.The mean patient age was 71 years  ml per session, which corresponds to 3 % polidocanol. The total treatment dose of sclerosant required for sufficient sclerotherapy averaged 16 (range 8–30) ml per patients.Table One patient (Case 2) had a fever of >38 °C and required an antipyretic after the second sclerotherapy session, but fever subsided the following day. Another patient (Case 3) experienced moderate pain at the puncture site for several days, which was relieved by intravenous administration of an analgesic. No patient experienced no life-threatening complications.2 are used commonly to treat varicose veins in the extremities because several randomized trials have shown higher efficacy rates and lower recurrence rates with foam than with liquids [Foam formulations of polidocanol mixed with room air or CO liquids , 10. One liquids . Polidoc liquids , but foaPolidocanol foam sclerotherapy has several potential advantages over conventional liquid sclerotherapy , 17: (1)Foam sclerotherapy has been indicated recently for vascular malformations , 12, andPolidocanol foam sclerotherapy is more cost-effective than 5 % EOI sclerotherapy. There is no consensus on the maximum dose of 3 % polidocanol suitable for daily foam sclerotherapy sessions for hepatic cysts. Ten milliliters of 3 % polidocanol was tentatively determined as the maximum dose per session in the present study, because the manufacturer recommends that the maximum daily dose should not exceed 30 ml of 1 % polidocanol for endoscopic injection sclerotherapy to treat esophageal varices. The net dose of 3 % polidocanol per patient averaged 16 (range 8–30) ml in the present study. Thus, sufficient sclerotic effects were achieved with a lower dose of sclerotic agent than was the case for the two previous studies of EOI sclerotherapy, in which the treatment session dose of 5 % EOI was fixed to 10 % of the initial cyst volume and the mean amount of 5 % EOI required for sclerotherapy was 30.9 (range 6–60) ml , respect2 is useful for predicting the behavior of the injected gaseous sclerosant and can confirm that no sclerosant has entered the systemic circulation or adjacent hepatic vessels. Injected sclerosant is easily identified by C-arm CT after injection, as the air density area replaces fluid within the cyst. This also confirms the absence of migration into the surrounding hepatic vessels. One case of polidocanol foam injection-associated moderate fever was observed in this study, which is not much higher than that reported with EOI sclerotherapy [In the phlebology field, reversible cardiac arrest  and tranotherapy , 7. Delaotherapy , was notCompletion of sclerotherapy over a single session is ideal, considering that long-term implantation of a catheter increases the risk of infection and discomfort. The mean number of treatment sessions for sufficient sclerotherapy in this study was two sessions. Our final case (Case 3) was successfully treated with only a single session despite having a cyst with a relatively large initial volume . This suggests that our first two cases may have been resolved with fewer sessions.The study limitations were (1) its single-arm retrospective design with a small number of patients from a single institute without any randomization and (2) the relatively short follow-up period.2 is a safe and effective treatment for symptomatic hepatic cysts that provides sufficient cyst volume reduction without severe complications. Further evaluation to determine the optimal dose and timing of polidocanol foam sclerotherapy relative to cyst volume are warranted, as are randomized controlled studies with long-term follow-up comparisons with EOI and sodium tetradecyl-sulfate foam.C-arm CT-assisted polidocanol foam sclerotherapy following digital subtraction cystography with CO


Low birth weight (LBW) is closely associated with foetal and neonatal mortality and morbidity, inhibite growth and cognitive development and resulted chronic diseases later in life. Many factors affect foetal growth and thus, the birth weight. These factors operate to various extents in different environments and cultures. The prevalence of low birth weight in the study area is the highest in the country. To the investigator’s knowledge in Bale Zone, no study has yet been done to elucidate the risk factors for low birth weight using case control study design. This study was aimed to identify the risk factors of low birth weight in Bale zone hospitals.A case–control study design was applied from April 1st to August 30th, 2013. A total of 387 mothers (136 cases and 272 controls) were interviewed using structured and pretested questionnaire by trained data collectors working in delivery ward. For each case, two consecutive controls were included in the study. All cases and controls were mothers with singleton birth, full term babies, no diabetes mellitus and no hypertensive. The data were entered and analyzed using SPSS version 16.0 statistical package. The association between the independent variables and dependent variable (birth weight) was evaluated through bivariate and multiple logistic regression analyses.Maternal age at delivery <20 years (adjusted odds ratio (AOR) = 3; 95 % confidence interval (CI) = 1.65–5.73), monthly income <26 United States Dollarr (USD) , lack of formal education , being merchant  and residing in rural area  were socio-economic variables associated with low birth weight. Maternal risk factors like occurrence of health problems during pregnancy , maternal body mass index <18 kg/m2 , maternal height <1.5m , inter-pregnancy interval <2 years  and history of khat chewing  and environmental factors such as using firewood for cooking , using kerosene for cooking , wash hands with water only  and not having separate kitchen room  were associated with low birth weight.Women who residing in rural area, faced health problems during current pregnancy, had no antenatal care follow-up and use firewood as energy source were found to be more likely to give low birth weight babies. Improving a mother’s awareness and practice for a healthy pregnancy needs to be emphasized to reverse LBW related problems. Low birth weight (LBW) is considered as the single most important predictor of infant mortality, especially of deaths within the first month of life . It is aMore than 20 million infants worldwide, representing 16 % of all births are born with low birth weight. The level of low birth weight in low income countries is more than double the level in middle income countries. About 10 % of births in Oceania were low birth weight births , 4. The The magnitude of LBW births are probably underestimates of the global situation because in the developing world a significant proportion of infants are born at home and not registered as live births . AccordiEthiopia is known to be among countries with very high maternal and child mortality rate. Even though sufficient specific data on Oromiya and Gambela are lacking, It would not be a stretch to assume that the grim statistics would apply to the women in the two regions too .Many factors determine the duration of gestation and foetal growth, and thus, the birth weight. They might be related to the infant, the mother, or the physical environment and play an important role in determining the birth weight and the future health of the infant . In diffBirth weight is affected to a great extent by the mother’s own foetal growth and her diet from birth to pregnancy. Mother’s poor nutrition and health, high prevalence of specific and non-specific infections, pregnancy complications, and physically demanding work during pregnancy are contributes to poor foetal growth .In order to prevent LBW, its main modifiable risk factors need to be understood. Additionally, the interrelationships between maternal, social and cultural factors need to be investigated. Results of the research would be critical to develop interventions aimed at modifying behaviors and other risk factors for low birth weight. Hence, this research was aimed to identifying the socio-economic, maternal and environmental risk factors for low birth weight in the study area to design urgent and sustainable interventions.st to August 30th, 2013. Bale zone is the second largest zone in Oromia regional state located in the South-eastern part of Ethiopia. The zone administratively divided in to 17 districts and 6 town administration [Bale Zone administrative office 2013]. Based on bale zone health office report, there are four government hospitals  and 76 functional health centers, 351 functional health post, 179 private clinic, 1 NGO clinic, 4 other public clinic, 95 pharmacy/drug shop, 1 NGO drug shop and 4 medical drug store in Bale zone.A hospital based case control study was conducted in Bale zone from April 1All mothers who gave birth in the four governmental hospitals were the source population. Mothers who gave live births weighed less than 2500 g were considered as cases and live births weighed 2500 g and above as controls. Mothers who had diabetes mellitus, hypertension, preterm baby and multiple births were excluded; because those conditions are known risk factors for low birth weight.α/2 = 1.96), 80 % power (Zβ = 0.84), control to case ratio 1:2 (r = 2), the odds ratio to be detected ≥ 2 and the 20 % control group will be exposed. The final sample size was 408 (136 cases and 272 controls).The sample size was determined using the proportion difference approach with the assumption of 95 % confidence level  and behavioral surveillance survey (BSS) and other peer reviewed articles , 6, 7. TThe weight of the newborns was measured within 15 min after birth using a balanced Seca scale. The scale was always checked and zeroed before weighing each newborn. Maternal height was measured against a wall height scale to the nearest centimeter. Maternal weight was measured by beam balance to the nearest kilogram and body mass index (BMI) was subsequently calculated.The first weight of the new-borns measured within 15 min after birth. Low birth weight (cases) were those newborns weighed less than 2500g while those newborns with birth weight of 2500g and above were considered as normal weight (controls).It is a birth before a gestational age of 37 complete weeks.It refers when more than one fetus is carried to term in a single pregnancy.First the data were checked for completeness and inconsistencies. Then coded and entered to SPSS version 16.0 soft ware. The entered data were cleaned and edited before subsequent analysis. Summary statistics such as mean and standard deviation was computed for cases and controls groups. The socio-demographic characteristics of the mothers were cross tabulated among cases and controls. Bivariate and multiple logistic regression analyses were done to identify the relationship between the independent variables  and dependent variable (birth weight).The socio-economic factors; maternal age, residence, marital status, maternal education, maternal occupation, husband’s occupation, husband’s education, monthly income and role of decision making on money how to be used were entered to the bivariate model with low birth weight. Similarly, maternal factors including; birth interval, gravida, antenatal care (ANC) follow-up, gestational age at first ANC visit, deworming during pregnancy, maternal height, maternal weight, maternal BMI, history of pregnancy related problems, history of alcohol drinking and khat chewing were entered in to the bivariate model. Likewise; environmental factors entered to bivariate analysis were latrine availability, average daily household water consumption, mothers’ hand washing practice, availability of separate kitchen room, source of drinking water, solid waste disposal site and water source point accessibility to household.The three sets of independent variables  that showed significant association in the bivariate logistic regression analysis were entered in multiple logistic regressions analysis using backward stepwise method. All statistical tests were two sided and significant association was declared at p-value less than 0.05.Ethical clearance letter was obtained from research review committee (ERC) of Madawalabu University. Permission letters were secured from Bale Zone Health Bureau and from the four respective hospitals. Verbal consent was obtained from each mother prior to interview. Additionally, all the information obtained from each study participant was kept confidential throughout the process of this study.From a total of 408 sample size, 387 mothers of (129 cases and 258 controls) were included in the interviewe which made the response rate of 94 % for both cases and controls.Almost half of mothers of the cases 51.2 % and more than two third of mothers of controls 69.4 % were in the age group of 21–35 years. About sixty-seven percent of mothers among cases and 53.9 % of mothers among controls were Muslim in religion. Larger proportions, 69.8 % of cases of mothers and 45.3 % of the controls mothers were housewives. About forty-six percent of mothers of LBW babies were illiterate while 15.5 % of mothers of normal birth weight (NBW) babies were illiterate. Concerning monthly family income, relatively higher percentage of mothers of low birth weight babies 24.6 % had an income less than 26$ ompared to mothers of normal birth weight babies 7.8 % [Table 2. Maternal height, 84.5 % from cases and 93.8 % from controls were greater than 150cm tall. Among mothers of cases 48.1 % and mothers of controls 24.8 % lived in rural part of the study area and most of mothers 93 % were currently married [Table Nearly half of mothers with LBW babies 50.8 % spaced between present and past pregnancy more than 2 years compared to mothers with NBW babies 74.7 %. Seventy six percent of mothers among cases and 82 % of mothers among controls had BMI of 18.5–25 kg/mBivariate logistic regression analyses were performed between socio-economic factors of mothers and low birth weight. The analyses revealed that maternal age, residence, maternal education, maternal occupation, husband’s occupation, husband’s education, monthly income and participation on decision on how money be used were statistically significant with low birth weight in the bivariate model. Those socio economic factors of the mothers which have significant association with low birth weight in the bivariate model were entered to multiple logistic regression analyses. The results showed that mothers who were residing in rural areas were two times more prone to deliver LBW babies than their urban counterparts ). Those mothers with monthly income less than 26$ were four times more likely to give LBW baby as compared to mothers with monthly income of greater than 79 $ ). Mothers who had no formal education were at higher risk to give low birth weight baby as compared to mothers with tertiary level of education ). Mothers who were in the age group of less than 20 years were more likely to deliver low birth weight babies than those mothers in the age group of 21–35 years ). Mothers who were merchant by their occupational were 90 % less likely to deliver low birth weight babies compared to employed mothers ) . Maternal short stature (≤150 cm) AOR = 3.7; 95 % CI = 1.22–11.28) and khat chewing  were risk factors for low birth weight [Table Similarly, bivariate logistic regression analyses were done to check the presence of significant association between maternal factors and low birth weight. As a result; birth interval, gravida, ANC follow-up, gestational age at first ANC visit, maternal height, maternal weight, maternal BMI, history of pregnancy related problems, history of alcohol drinking and history of khat chewing were statistically associated with low birth weight. In multiple logistic regression analysis; mothers who encountered pregnancy related health problems during current pregnancy were at higher risk to deliver low birth weight baby than mothers who didn’t encounter any health problem . The odds of low birth weight were higher among mothers who didn’t attend antenatal care for current pregnancy as compared to mothers who attended ANC . In the same manner; mothers with birth interval of 2 years and below between the current and previous birth were more likely to give low birth weight baby than mothers who gave birth greater than 2 years apart ). The odds of giving LBW baby were higher among mothers with body mass index (BMI) less than 18.50kg/mThe household environmental factors including latrine availability, average daily household water consumption, and mothers’ hand washing practice, availability of separate kitchen room, solid waste disposal site and source of energy for cooking were statistically associated with LBW in the bivariate logistic regression analyses model. The multiple logistic regression results showed that; the likelihood of giving low birth weight baby was significantly higher among mothers who were used firewood for cooking than electricity , kerosene than electricity  and animal dung than electricity ). Mothers from household which had no separate room for cooking significantly associated with low birth weight . Mothers who washed their hands with water only had higher probability of giving low birth weight baby than mothers who washed their hands using water with soap  and hand washing with water and ash also found to be risky for low birth weight compared to using water with soap . The odds of LBW babies among mothers with daily household water consumption less than 50 l were higher than mothers with daily household water consumption of 50 l and above  [Table Low birth weight can be influenced by various factors that occur prior to and during pregnancy including the household environmental conditions where the mothers live. Therefore; this study identified the risk factors for low birth weight which is important for proper, immediate and sustainable intervention to improve maternal health for better pregnancy outcome ,11.This study showed that some of the socio-economic conditions affect the weight of new born negatively. In this regard, mothers who resided in rural areas were more likely to deliver low birth weight babies. This finding is in agreement with study done in Tanzania and India , 13. ButThis study revealed that mothers who are illiterate and in lower income level were at higher risk to deliver LBW babies. Similarly, the study conducted in Nepal and Lahore showed that maternal education and per capita income of the family per month were found to be significantly associated with birth weight of the new born , 15. TheHowever, this study revealed no association between occupational status and LBW and lack of decision power on their resource utilization and LBW which is different from other previous study findings , 16. ThiPregnancy is a life threatening condition in a majority of developing countries, Its anomalous outcome reduces the life expectancy of new borns and their mothers. In this study, mothers who encountered pregnancy related health problems during current pregnancy were at higher risk to deliver low birth weight baby than mothers who didn’t. This result is similar with a study done in India that showed mothers with any health problem during pregnancy were two times more likely to give low birth weight babies .The risk of low birth weight was higher among mothers who didn’t attend antenatal care for current pregnancy as compared to mothers who attended ANC. This is consistent with a study done in Nepal which showed as birth weight was significantly associated ANC service utilization .Antenatal visits of the pregnant mothers are very important as they provide chances for monitoring the fetal wellbeing and allow timely intervention for feto-maternal protection including nutritional counseling that a mother might receive. Likewise, birth spacing had significant association with LBW. Mothers with birth spacing of 2 years and below were more likely to deliver low birth weight baby than mothers who delivered with birth interval of 2 or more years. This finding is in-line with a study done in India that showed birth interval of < 2 years were at higher risk to deliver LBW baby . These f2 and height less than 1.50 m were more likely to deliver low birth weight babies. This findings were consistent with a study conducted in India which revealed that low birth weights were significantly higher among mothers with height <145 cm and BMI <18.5 kg/m2 [We found that mothers BMI less than 18 kg/m.5 kg/m2 . It is a.5 kg/m2 , 18.2 were significantly higher in mothers who had LBW babies compared to those who delivered NBW babies in another case control study in Iran [It is also consistent with another similar study where BMI (<18 kg/m2) two times prone to deliver low birth weight babies . The mea in Iran . This miIn this study maternal age at first birth, history of alcohol drink and number of pregnancies didn’t have significant associations with low birth weight. But mothers who had history of Khat chewing were statistically higher at risk to deliver LBW as compared to mothers who didn’t chew Khat.The sources of drinking water affect the health of the people that use it. If toilet facilities, water sources and cooking environment are poor among mothers, it will expose them to various infections that leads to poor pregnancy outcomes. Various household environmental factors have been implicated in adverse pregnancy outcomes. The combustion product of solid fuel in developing countries can cause many adverse health effects in people. Majority of pregnant women in developing countries are heavily exposed to indoor air pollution which attributes to low birth weight . This stIn this study, mothers who didn’t have separate room for cooking were more likely to experience low birth weight babies. This could be due to maternal exposure during pregnant to smoky kitchens which is not separated from the dwelling room might result to inhale chemicals from biomass fuels which contribute for low birth weight and perinatal mortality , 27.The findings of this study showed that the presence of significant association between the socio-economic, maternal and household environmental factors and birth weight of the new-borns among mothers who gave birth in Bale zone hospitals.From socio-economic factors; not having formal education, being a resident in rural area, maternal age less than 20 at current birth and having monthly income less than 26$ were identified as risk factors for low birth weight.2, presence of pregnancy induced health problems and having history of Khat chewing were among maternal factors identified as positively associated with low birth weight.Absence of antenatal care follow-up, birth spacing of 2 years and below, short maternal stature, maternal BMI of less than 18Kg/mNot having separate room for cooking, being firewood user, being kerosene user & being animal dung user as energy source for cooking, and lack of proper hand washing practice such as use of water only or water with ash rather than water with soap were among the household environmental conditions that increase the risk of low birth weight baby.This study identified various socio economic, maternal and environmental risk factors for low birth weight. Therefore; prevention strategy for low birth weight in this area should be designed to tackle these multiple risk factors for low birth weight. Income generation means such as small scale enterprises should give due attention for mothers. In addition; mothers should be encouraged to use family planning method so as to maximize birth intervals between subsequent births.Health professionals should screen and consulate pregnant mothers who are at risk of having infants with LBW and ensure that women have access to essential health information on the causes of low birth weight. Public education and awareness on how to carry on a healthy pregnancy. Likewise; women should be linked to the appropriate maternal health services including antenatal care and nutritional counseling services.Community sensitization should do to improve household environmental conditions, where the pregnant women live and work. This should be in focus of promoting to have separate kitchen from living rooms and to use non-smoky energy sources for cooking such as electricity or to be away from such activities.


The purpose of this study was to assess the impact of maternal DM on fetal adipokines and anthropometry in infants of Hispanic and Native American women.Offspring of women with diabetes mellitus (DM) during pregnancy have a risk of developing metabolic disease in adulthood greater than that conferred by genetics alone. The mechanisms responsible are unknown, but likely involve fetal exposure to the n = 79) or type 2 or gestational DM  pregnancies. Infant anthropometrics were measured at birth and 1-month of age. Cord leptin, high-molecular-weight adiponectin (HMWA), pigment epithelium-derived factor (PEDF) and C-peptide were measured by ELISA. Differences between groups were assessed using the Generalized Linear Model framework. Correlations were calculated as standardized regression coefficients and adjusted for significant covariates.We conducted a prospective study of offspring of mothers with normoglycemia , but sum of skinfolds (SSF) were not different. At 1-month, there was no difference in weight, SSF or % body fat or postnatal growth between groups. Leptin was higher in DM-O . Leptin was positively associated with birth weight (p = 0.0007) and SSF (p = 0.002) in Con-O and with maternal hemoglobin A1c in both groups . PEDF was positively associated with birth weight in all infants (p = 0.004). Leptin was positively associated with PEDF in both groups, with a stronger correlation in DM-O (p = 0.009). At 1-month, HMWA was positively associated with body weight (p = 0.004), SSF (p = 0.025) and % body fat (p = 0.004) across the cohort.DM-O were heavier at birth than Con-O (3.7 ± 0.6 vs. 3.4 ± 0.4 kg, in utero exposure to DM imparts hormonal differences on infants even without aberrant growth.Maternal DM results in fetal hyperleptinemia independent of adiposity. HMWA appears to influence postnatal growth. Thus,  First, the risk associated with maternal DM is greater than that of paternal DM. Second, fetal exposure to the maternal diabetic milieu confers additional risk, independent of other factors. Dabelea and colleagues reported that offspring conceived after their mother’s DM diagnosis are nearly four times more likely to develop T2DM than their siblings born before their mother developed diabetes, supporting a key role for the fetal environment [Children born to mothers with diabetes mellitus (DM) are at increased risk for development of type 2 diabetes mellitus (T2DM) and other metabolic abnormalities later in life. The mechanisms of transmission to offspring are not yet understood and go beyond straightforward Mendelian inheritance of susceptibility genes. Several lines of evidence implicate the ironment .in utero transmission of risk for diabetes from a mother to her offspring, whether genetic or environmental, potentially involves modulation of circulating cytokines that influence appetite regulation, insulin action, or other metabolic functions. We selected leptin and high-molecular-weight adiponectin (HMWA) for evaluation because both are produced by adipose tissue and have been shown in studies of adults or experimental animals to be associated with insulin resistance, adiposity, fat metabolism, and inflammation.The Pigment epithelial-derived factor (PEDF) is a serpin-like protein lacking inhibitory function that was selected for study because expression is known to be influenced by diabetes and obesity , 3, circin utero milieu) and anthropometric measures at birth and at 1-month of age in offspring of Hispanic and Native American women, a population at intrinsically increased risk for the development of T2DM [We investigated the impact of maternal DM on the concentrations of these adipokines in fetal circulation , the Chickasaw Nation Medical Center  and the Choctaw Nation Health Care Center  between April 2010 and October 2013. Signed informed consent was obtained from a parent of each infant in accordance with the University of Oklahoma Health Sciences Center, Chickasaw Nation and/or Choctaw Nation of Oklahoma Institutional Review Boards, each of which approved the study.in utero exposure to DM on postnatal growth. Maternal gestational and type 2 diabetes were diagnosed according to ADA guidelines [Offspring of self-identified Native American and Hispanic women with diagnosed gestational or pre-existing type 2 diabetes mellitus (DM-O group) or confirmed normoglycemic pregnancies (Con-O group) were enrolled at birth into a prospective, longitudinal study of the impact of idelines . InfantsMaternal age at delivery, gestational weight gain, hemoglobin A1c (if available), results of glucose tolerance screenings, pregnancy history, smoking status and any medications taken during pregnancy were extracted from medical records. Maternal height and pre-pregnancy weight were obtained from medical records if recorded, or by self-report if not, and used to calculate pre-pregnancy body mass index (BMI).At 1-month of age, infants visited the CMRI Metabolic Research Program facilities at the University of Oklahoma Health Sciences Center for follow-up measurements. Length, weight, head and abdominal circumferences and skinfold thickness were measured and body composition was assessed by dual energy x-ray absorptiometry . All DXA scans were performed and analyzed by the same person (DAF); suitable scans were obtained in 98.4 % of the infants.Leptin, high-molecular-weight adiponectin (HMWA), pigment epithelium-derived factor (PEDF) and C-peptide concentrations were measured by ELISA  according to the manufacturer’s protocol. All samples were assayed in a single run. Intra-assay variability for leptin, HMWA, PEDF and C-peptide were 2.1 %, 3.2 %, 6.9 %, and 5.8 %, respectively.p = 0.010; no difference in the DM-O group). Analyses at 1-month include data on all infants for whom both the birth and 1-month visits were completed . Participants who completed only the birth visit were not different from those who completed both visits in terms of gestational age, birth anthropometrics or maternal characteristics. The proportions of infants fed formula and/or breastmilk were similar between the groups (% breastmilk/formula/mixed were 24.4/55.6/20.0 in Controls and 20.6/52.9/26.5 in DM). Primary outcomes were infant weight and length; skinfold thickness; percent body fat; fat, lean and fat-free mass; cord serum leptin, HMWA, PEDF and C-peptide.Birth analyses include all mother-infant pairs for whom umbilical cord blood, anthropometric measures and maternal records  were available. Thus, data for 79 out of 94 controls and 45 out of 65 DMs were included. Subjects who had cord blood samples available were not different from those who did not in terms of birth anthropometrics or maternal characteristics, except that gestational age was slightly less in those without cord blood samples in the Con-O group , and maternal pre-pregnancy BMI. The covariates included in the model in each case were selected using stepwise methods. While cord blood HMWA and C-peptide values were normally distributed, leptin and PEDF had skewed sampling distributions and were logarithmically transformed to meet the assumptions for linear regression analysis. Correlations were calculated as standardized regression coefficients and adjusted for significant covariates. Data analyses used IBM SPSS Statistics . P-values <0.05 were treated as significant for the purposes of discussion.2) or obese (BMI ≥ 30.0 kg/m2) by Centers for Disease Control criteria (median BMI = 27.6) whereas 90 % of DM mothers were overweight or obese (median BMI = 32.1). Maternal weight gain during pregnancy for the DM group was about 3 kg less than that of control mothers, but not significant. Parity was higher for DM mothers than for control mothers, possibly reflecting their older average age.Mothers with DM were 6.4 years older on average and reported increased parity, pre-pregnancy weight and BMI compared to control mothers Table . Fifty-sWomen with gestational diabetes comprised the majority of the DM group (32 GDM vs. 13 T2DM). There was no difference in cord blood adipokines in offspring of women with T2DM versus GDM so we combined them for subsequent analyses. Dysglycemia generally was well controlled (Table p = 0.024), though length and SSF were equivalent between the groups.All infants were born at term , but this was no longer a statistically significant difference. There also were no between-group differences in length or SSF and no differences in % fat in the whole body, trunk or limbs as measured by DXA. Furthermore, the change in weight, length and SSF from birth to 1-month did not differ between DM-O and Con-O  with DM-O showing a stronger correlation. Cord PEDF also correlated significantly with maternal HbA1c when the groups were combined (p = 0.047).Only seven of the 40 mothers for whom data were available had an HbA1c greater than 6 %. Significant correlations involving HbA1c remained significant with and without those seven subjects; therefore we chose to include them in the analyses. Maternal HbA1c correlated positively with cord leptin level in the entire cohort Table  and withIn contrast with leptin, there was no effect of DM status on cord blood concentrations of HMWA or C-peptide.Examination of the relationship between leptin and birth anthropometrics Table  revealedp = 0.0042) with cord concentration  was unrelated to the concentration of cord leptin in infants of DM mothers, an effect that was independent of birth weight and gestational age. Several previous publications describe increased cord blood leptin levels in offspring of women with T2DM, T1DM, or GDM. While there is general agreement that maternal DM raises cord blood leptin level  for the first six months of life , supportin vitro [The role of PEDF during human development and childhood is largely unexplored. We found a modest relationship between birth weight and cord blood PEDF overall with no differences between infants born to mothers with DM versus controls. However, there were striking correlations with cord blood levels of both HMWA (negative) and leptin (positive). The strongest relationship was between leptin and PEDF in the offspring of DM mothers, wherein 38 % of the variation in leptin concentration was explained by PEDF level. Although the basis for this relationship is unclear, it could be reflect the need for the angiogenic properties of leptin to be balanced by the anti-angiogenic effects of PEDF in the placenta, a putative source of cord blood leptin and PEDF , 29. PEDin vitro . Furtherin vitro , providiThe strengths of this study are the careful phenotyping of the infants at birth, the detailed measures of growth and body composition over the first month of life, and the new insights concerning potential determinants of cord leptin, PEDF, and the connection between HMWA and the acquisition of adipose tissue. The conclusions, however, are based on correlations and thus cause and effect relationships remain to be determined. In addition, we do not know whether the observations concerning PEDF and leptin reflect changes in expression by placenta or other fetal tissues such as liver or adipose depots. A potential limitation is the attrition between the birth and 1-month visits . However, there were no obvious distinctions between subjects who completed only the birth visit and those who completed both visits, suggesting this had minimal impact on our findings. Treatment modalities varied amongst the DM mothers, with glyburide and insulin most commonly prescribed. Though neither birth weight  nor neonCord blood leptin at term is increased by maternal diabetes independently of neonatal adiposity and is directly related to maternal hemoglobin A1c percent in offspring of diabetic and non-diabetic women. These data provide further evidence that infants born to mothers with near-optimal management of diabetes still show the effects of the condition and that the level of maternal glycemia has a major influence on leptin secretion. In contrast, PEDF concentration is unaffected by dysglycemia, but correlates with birth weight and leptin, suggesting a novel role(s) for this understudied cytokine in fetal growth control and placental function. Lastly, higher levels of cord blood HMWA were associated with increased infant growth and adiposity at 1 month, supporting a role for HMWA in the acquisition of fat mass during infancy.


To investigate the effect of parental personality on birth outcomes.Prospective cohort study.727 pregnant women and 579 spouses receiving antenatal care at a single-center in rural Tokyo, Japan during 2010–2013.We measured the association between maternal effect of parental personality traits assessed by the Cloninger’s Temperament and Character Inventory on birth outcomes, using multiple regression and adjusting for demographics.Maternal self-transcendence personality was inversely associated with gestational age [-0.26 : -0.51 to -0.01) weeks per unit] and positively associated with preterm birth [odds ratio (OR) 2.60 (95% CI: 1.00 to 6.75) per unit], while paternal self-transcendence personality was positively associated with gestational age [0.31 (95% CI: 0.07 to 0.55) weeks per unit]. Maternal reward dependence was positively associated with fetal growth [0.30 (95% CI: 0.02 to 0.59) per unit]. Other maternal and paternal personality traits associated with adverse maternal behavior, such as novelty seeking, harm avoidance and self-directedness, were not associated with birth outcomes.We found that specific parental personality traits can be associated with birth outcomes. Preterm birth and restricted fetal growth have long been the most significant parameters of adverse outcomes in maternal-child health in many countries. Increasing knowledge has shown that their effects on children’s health are not limited to higher morbidity risk,  but longRecent research has shown that various maternal behaviors such as smoking ,10, inadPrevious studies measuring behavior using the Temperament and Character Inventory , a widelHowever, no study using a valid personality assessment scale, including the two above, has assessed how birth outcomes are affected by maternal and paternal personality. Therefore, we examined the association between antenatal maternal and paternal personality and birth outcomes, such as preterm or small for gestational age (SGA), using the Temperament and Character Inventory .This study protocol was approved by the Institutional Review Board at the National Center for Child Health and Development on August 2, 2010 (Study ID: H22-417). All participants provided written informed consent to participate in this study. Our study was done using a sub-cohort of the Seiiku Boshi Cohort, a prospective cohort study on pregnant women, their spouses and children. For this cohort, pregnant women attending antenatal visits at the National Center for Child Health and Development (NCCHD) from May 13, 2010 until Nov 28, 2013 were recruited during their first antenatal visit, which usually takes place in weeks 6–14 of gestation . The NCCHD hospital is a tertiary hospital located in suburban Tokyo, and manages approximately 1,500 annual deliveries. Exclusion criteria were women whom nurses considered recruitment may be a burden, such as those diagnosed as threatened abortion. In total, 2,309 women consented to participate in this study. Participants were asked to fill out questionnaires, and medical records for the mother and child were retrieved from hospital charts for all deliveries at the hospital.Among these women, a sub-group of women with single pregnancies were approached during their regular antenatal visits to additionally asked to fill out the Temperament and Character Inventory in the second trimester (N = 727). Of this sub-cohort, 579 spouses also completed the Temperament and Character Inventory.We measured maternal and paternal personality in the second trimester using the Temperament and Character Inventory created by Cloninger ,25, whicCloninger categorized personality into two domains of temperament and character . ‘TemperWe collected maternal socio-demographic and other characteristic data and categorized them as follows: parental age, parity , annual household income , maternal employment status , maternal education , smoking during pregnancy , and daily exercise during pregnancy . We treated maternal and paternal age as continuous variables.Gestational age as well as birth weight, height, and head circumference were obtained from medical charts. Fetal growth was calculated as gestational age-adjusted birth weight z-scores, using Japanese birth weight curves for reference .First, we evaluated correlations among the seven personality traits for the participating women (i.e. four dimensions of temperament and three of character), as well as for the participating men. We also assessed cross-correlation between the personality traits of participating couples using Pearson’s correlation co-efficient.Second, we used multivariate linear regression to assess whether each maternal/paternal personality dimension score was associated with birth outcomes, adjusting for parental socio-demographic characteristics. As maternal personality dimensions are inter-correlated, and maternal and paternal personality dimensions are also correlated, we repeated this analysis including all maternal characteristic traits in one model. We did not adjust for maternal complications, behaviors or mental disorders as we considered these caused by personality and would be mediators than confounders. All statistical analyses were conducted using the statistical software package Stata 13 , and the p-value <0.05 was considered as statistically significant when performing hypothesis tests.In Similarities could be seen among the personality traits of expectant mothers and fathers, with maternal and paternal harm avoidance, reward dependence, self-directedness, cooperativeness and self-transcendence having a significant weak correlation. Among the participating couples, one parent’s reward dependence was weakly but significantly correlated with the cooperativeness of the other, and the harm avoidance of one parent was weakly but significantly negatively correlated with the self-directedness of the other. In addition, maternal persistence and paternal self-transcendence were weakly negatively correlated.In Tables Similarly, paternal self-transcendence was significantly associated with higher gestational age [0.31 (95% CI: 0.07 to 0.55) weeks per unit] but not risk of preterm birth or SGA, with the association persisting after adjustment for other maternal and paternal characteristics, including other personality traits. Other paternal personality traits were not associated with birth outcomes.We investigated the association between maternal and paternal personality measured by the Temperament and Character Inventory, and birth outcomes. We found several traits of antenatal maternal and paternal personality to be a predictor of birth outcomes, especially gestational length. Higher maternal self-transcendence was associated with shorter gestational length and higher risk of preterm birth; higher maternal reward dependence was associated with greater fetal growth, and higher paternal self-transcendence was associated with longer gestational length. However, personality traits including novelty seeking, self-directedness, harm avoidance or cooperativeness associated with adverse behaviors (smoking) or mood disorders (depression) did not show significant association with neither fetal growth nor gestational length.Personality is an important factor for how people behave. As maternal and paternal behavior is likely to affect the fetus, we hypothesized that parental temperament would be associated with birth outcomes. However, the associations between temperament and birth outcomes that we observed could not be explained by known behaviors, such as smoking.As for the association between smoking and maternal temperament, we found that novelty seeking, which is known to be associated with a decrease in basal dopamine and an increase in cluster B (impulsive) personality disorders , was assAnte-natal depression could also be a mediating factor between personality and birth outcomes, as higher harm avoidance ,22 and lSelf-transcendence, a character dimension of personality, is described as the experience of spiritual concepts; for example, regarding oneself as being at one with the universe . People Reward dependence, an inheritable temperament dimension of personality, relates to social sensitivity and dependence on the approval of others . People Our study has several limitations. First, as the study was conducted in an area of Japan with a fairly well-educated, relatively higher socioeconomic-status population, our findings may not be generalizable to other cultural backgrounds. However, some of our results did show similarities with previous studies, such as the correlation among personality traits of one individual ,29. SecoWe found specific parental personality traits can be associated with birth outcomes, however failed to explain these observed relationships by behaviors associated with personality. Further research to elucidate how parental personality influence birth outcomes by altering behavior is needed.