Characterization of an actin-binding site within the talin FERM domain

Talin is a large cytoskeletal protein that couples integrins to F-actin. Three actin-binding sites (ABS1-3) have been reported: one in the N-terminal head, and two in the C-terminal rod domain. Although the C-terminal ABS3 has been partially characterized, the presence and properties of ABS1 within the talin head are less well defined. We show here that the talin head binds F-actin in vitro and in vivo at a specific site within the actin filament. Thus, purified talin head liberated from gizzard talin by calpain cleavage cosediments with F-actin in a low salt buffer at pH 6.4 (conditions that are optimal for binding intact talin), and using recombinant polypeptides, we have mapped ABS1 to the FERM domain within the talin head. Both the F2 and F3 FERM subdomains contribute to binding, and EGFP-tagged FERM subdomains colocalize with actin stress fibers when expressed in COS cells. High-resolution electron microscopy of actin filaments decorated with F2F3 localizes binding to a site that is distinct from that recognized by members of the calponin-homology superfamily. Finally, we show that the FERM domain can couple F-actin to PIPkin, and by inference to integrins, since they bind to the same pocket in the F3 subdomain. This suggests that the talin FERM domain functions as a linker between PIPkin or integrins and F-actin at sites of cell-matrix adhesions.     

Dystocia following prolonged retention of a dead fetus in an Asian elephant (Elephas maximus)

A 32-year-old nulliparous female Asian elephant (Elephas maximus) showed signs of parturition 8 months later than predicted from the breeding records. However, while serosanguineous fluid, necrotic tissue and pieces of amnion were expelled, second-stage labor did not progress. Since the fetus was not found during an endoscopic examination of the vestibule, it was assumed that the elephant had calved unseen and she was returned to the forest to recuperate. Twelve months later, the elephant showed clear signs of second-stage labor accompanied by a bulge in the perineum and passage of keratinized nail through the vulva. A 35 cm episiotomy incision was made in the perineum just below the anus, via which chains were attached to the forelimbs of the fetus. Traction on the forelimbs alone proved insufficient to achieve delivery because the fetal head kept rotating and impacting in the pelvis. However, traction applied via a hook inserted behind the mandibular symphysis allowed the head to be elevated and extended, and the fetus to be delivered. The episiotomy wound was sutured in two layers and although the skin did not heal during primary closure it subsequently healed uneventfully by second intention. Retrospective evaluation of the elephant's serum progestagens profile demonstrated a fall to baseline at the suspected onset of parturition, supporting the supposition that the fetus was retained in the uterus for 12 months after parturition began. It is suggested that serum progestagens concentrations should be monitored regularly in mated elephant cows to verify the establishment of pregnancy and to better estimate the expected timing, and the onset of calving.     

Evaluation of a Semiautomated System for Direct Fluorescent Antibody Detection of Salmonellae

A semi-automatic system under development by Aerojet Medical and Biological Systems for the direct fluorescent antibody detection of salmonellae was evaluated with various food, feed, and environmental samples. All samples were simultaneously examined by Automated Bioassay System (ABS), manual direct fluorescent antibody procedures and cultural procedures. The ABS gave satisfactory results with the processed samples. It detected all of the culturally positive powdered egg and candy samples with no false negative results and gave only 6.6 and 5.3% false positive rates, respectively. With meatmeal samples the ABS failed to detect one culturally positive specimen that was also positive by manual fluorescent antibody and gave one (1.1%) false-positive result. A high rate of false-negative results was obtained by ABS on unprocessed samples of creek water, poultry, and sausage. Adding another enrichment step to the protocol reduced the false-negative rate considerably but severely increased the false-positive rate. The instruments worked reasonably well, but research is needed to improve enrichment procedures for samples to be processed by the system.     

Amniogenic band anomalies in a fifth-month fetus and in a newborn from maternal oophorectomy during early pregnancy

A fifth-month fetus and a newborn with amniogenic band anomalies were examined at autopsy. Both specimens were obtained from women who had undergone oophorectomy during early pregnancy. The dead male fetus was aborted spontaneously, and had a micrognathia, a right club foot, and a constriction ring on the left lower leg. The left fingers 2, 3, and 4 were attached to the placenta by a fibrous string. No internal anomaly was noted. In the other case, a male newborn was delivered at the 39th week of gestation and had an agenesis of the calvarium, a cleft lip with palate, an amputation of the right toe, and constriction rings on right fingers 3 and 4 and left finger 3. The placenta was attached to the left temporooccipital region of the head by a fibrous string. Also present was an atrial septum defect and a horseshoe kidney. Possible etiology is discussed in relation to the "amniogenic bands" hypotheses.     

Spontaneous complete clefting of the palates in a mouse fetus: a study by scanning electron microscopy and serial section reconstruction

A 15 day mouse fetus having spontaneous complete clefting of the primary and secondary palates was studied in comparison with its normal litter mates and with normal 14 day fetuses. Specimens were studied by scanning electron microscopy at various stages of microdissection, by light microscopy of thin serial sections and by serial section reconstruction of the anterior chondrocranium of the clefted specimen and one of its normal litter mates. Differentiation of tooth and bone tissue was slightly retarded in the clefted fetus but paranasal and oral landmarks, though distorted, were present. The clefted fetus had a smaller angle between cranial base and nasal capsule and a marked discontinuity between the primary and secondary palates. Cell surfaces on the medial edge of the secondary palate in the clefted fetus resembled cell surfaces of oral areas that do not normally fuse, i.e. they are polygonial, flat and bear few surface projections in contrast to the normal 14 day condition where these cells are spindle shaped, convex and have many microvilli. The observations support the concepts that clefting of the secondary palate is consequential to clefting of the primary palate, that maldevelopment of neural crest mesenchyme is not necessarily a contributing factor, that clefting of the primary and secondary palates is associated with a shorter anterior-posterior dimension of the head and that when fusion of palatal shelves fails to occur the cells of the medial edges modulate in the direction of a generalized type of surface epithelium.     

Mobile shielding evaluation on the fetal dose during a breast radiotherapy using Monte Carlo simulation

PurposeEvaluation of the out-of-field dose is an important aspect in radiotherapy. Due to the fetus radiosensitivity, this evaluation becomes even more conclusive when the patient is pregnant. In this work, a linear accelerator Varian Clinac 2100c operating at 6 MV, a pregnant anthropomorphic phantom (Maria), and different shields added above the abdominal region of the phantom were used for the analysis based on MCNPX. Methods: The simulations were performed for the medial and lateral projections, using either an open field collimation (10×16 cm²) or a multileaf collimator. The added shields (M1 and M2) were designed based on models proposed by Stovall et al. [1], intending to reduce the deposited dose on the fetus and related structures. Results: The presence of the shields showed to be effective in reducing the doses on the fetus, amniotic sac, and placenta, for example. A reduction of about 43% was found in the dose on the fetus when M2 was added, using the open field collimation, in comparison with the situation with no shield, being the lateral projection the main responsible for the dose. The use of MLC significatively reduced the doses in different structures, including on the fetus and amniotic sac, for example, in comparison to the open field situation. A slight increment on the dose in organs such as the eyes, thyroid and brain was found in both collimation systems, due to the presence of the shields. The contribution of the leakage radiation from the tube head of the linear accelerator was found to be in the order of µGy, being reduced by the presence of the M2 shield. Conclusion: Using the shields showed to be an essential feature in order to reduce the dose not only on the fetus, but also in important structures responsible to its development.     

Delivering hydrocephalic fetuses

Detection of fetal hydrocephalus with head enlargement in the third trimester raises questions concerning the extent of the physician's obligations to the fetus and to the mother. Here Strong develops and defends an approach to these questions that he discussed in an earlier essay ("Ethical conflicts between mother and fetus in obstetrics," Clinics in Perinatology 1987 Jun; 14(2): 313-328), dividing the ethical issues involved into two main topics. He first explores under what circumstances a physician is ethically justified in draining fluid from the fetal cranium to reduce head size. This procedure, which usually causes fetal death, facilitates vaginal delivery. A cesarean section, which is less stressful for the fetus, exposes the woman to the risks of surgery. Secondly, Strong applies this discussion to the issue of how the physician should counsel the woman, and what recommendations, if any, the physician should make concerning the method of delivery.     

Solar UV-B effects on PSII performance in Betula nana are influenced by PAR level and reduced by EDU: results of a 3-year experiment in the High Arctic

The long-term and diurnal responses of photosystem II (PSII) performance to near-ambient UV-B radiation were investigated in High Arctic Betula nana. We conducted an UV exclusion experiment with five replicated blocks consisting of open control (no filter), photosynthetic active radiation and UV-B transparent filter control (Teflon), UV-B-absorbing filter (Mylar) and UV-AB-absorbing filter (Lexan). Ethylenediurea (EDU), a chemical normally used to protect plants against ozone injury, was sprayed on the leaves both in the field and in an additional laboratory study to investigate if EDU mitigated the effects of UV-B. Chlorophyll-a fluorescence induction curves were used for analysis of OJIP test parameters. Near-ambient UV-B radiation reduced across season maximum quantum yield (TR(o) /ABS = F(v) /F(m)), approximated number of active PSII reaction center (RC/ABS) and the performance index (PI(ABS)), despite improved leaf screening against UV-B with higher content of UV-B-absorbing compounds and a lower specific leaf area. EDU application counteracted the negative impact of UV-B on TR(o) /ABS, RC/ABS and PI(ABS) . This indicates that the mechanisms behind UV-B and ozone damage share some common features. The midday depression was present in all treatments, but TR(o) /ABS and PI(ABS) were persistently lower in near-ambient UV-B compared to UV-B reduction. The recovery phase was particularly impaired in near-ambient UV-B and interactive effects between treatment × hour raised TR(o) /ABS, RC/ABS and PI(ABS) higher in reduced UV-B compared to near-ambient UV-B. This demonstrates current solar UV-B to reduce the PSII performance both on a daily as well as a seasonal basis in this High Arctic species.     

Removal of ABS from solutions by a common fungus of sewage

Conclusions Phialophora jeanselmei (Margarinomyces heteromorphum) is very common in sewage treatment systems, and can remove ABS from solution even in the absence of glucose. Because glucose is present in sewage treatment systems either as a pollutant or as a byproduct of the metabolism of other organisms, it appears that, to some extent, ABS is removed by this organism. Additional work is needed to determine whether this organism can be made a nucleus of an artificial population deliberately established to act upon some special types of wastes; whether a mononucleate-monospore culture capable of more rapid and more complete removal of ABS can be developed; and what pathways of metabolism and decomposition of ABS might be involved with the idea of breaking into the chains of reaction with other organisms.In the presence of added glucose,Phialophora jeanselmei was found to be capable of removing ABS from shaken and still culture solutions containing an ABS concentration of 10 ppm. More adsorption on the mycelium took place in still culture than in aerated culture. In shaken culture a greater loss of ABS from solution resulted; this may be due to metabolic processes since the ABS was neither found in solution nor adsorbed on the mycelium. While wild cultures are capable of a demonstrable amount of activity, culture efficiency could be improved if selected monospore-mononucleate cultures were developed.     

Efficacy of Ankaferd Blood Stopper on bone healing in diabetic rats: a stereological and histopathological study

We evaluated the effects of Ankaferd Blood Stopper (ABS) and routine antibiotic prophylaxis (AP) on early healing of bone defects in diabetic rats. We used 48 rats in the study. Diabetes was induced in 24 rats using streptozotocin; the remaining 24 healthy untreated rats served as controls. Twelve of the diabetic rats and 12 of the healthy rats were treated with AP for 3 days before surgery. Bilateral bone defects were created in the mandible of all animals. ABS was applied to the defects on the left sides of the mandibles, while nothing was applied to the right sides. Animals were sacrificed on days 7 and 14 after operation and examined for histopathology and by stereology. The volume of newly formed bone was significantly less in the diabetic rats on both days 7 and 14. Local administration of ABS significantly increased the mean volume of newly formed bone in both diabetic and nondiabetic rats at days 7 and 14. No significant difference in new bone formation was found between AP and ABS treatment in diabetic rats. Both AP and local administration of ABS have beneficial effects on bone healing in diabetic animals.     

A new aqueous biphasic system containing polypropylene glycol and a water-miscible ionic liquid

In this work, we proposed a novel aqueous biphasic system (ABS) composed of polypropylene glycol P400 (PPG P400) and hydrophilic ionic liquids (IL), 1-alkyl-3-methylimidazolium bromide (alkyl = ethyl or butyl), forming an upper polymer-rich phase and a lower IL-rich phase at ambient temperature. This new ABS can present interesting characteristics shared by ILs and polymers such as low volatility, good solvation ability, tunable physical properties, and high design capacity for achieving task-specific phase components to enhance the partitioning of target species. Ternary phase diagram of the novel ABS formed by PPG 400 and [C(2) mim]Br in water was measured at T = 298.15 K. Factors affecting the binodal curves such as the cation side alkyl chain length and the temperature were also evaluated. The results were successfully interpreted in terms of the kosmotropic/chaotropic nature of ILs. Furthermore, the phase behavior of the PPG-[C(2) mim]Br ABS is described by the NRTL model. Finally, the extraction potential of the proposed ABS was evaluated through its application to the extraction of the essential amino acids such as L-tryptophan and L-tyrosine. The partition coefficients here obtained demonstrated the fine potential of the proposed ABS for biomolecules separation.     

Elimination from bone marrow of a supplementary population participating in the proliferation of pluripotent hematopoietic stem cells using mouse brain antiserum

Changes in the number of spleen exo-colonies and post-radiation repopulation of hematopoietic organs were studied in recipients upon injection of bone marrow treated with anti-brain serum (ABS) with and without thymocytes on days 9-14. It was shown that on days 9-11 colony formation in mice injected bone marrow treated with ABS was much lower than the control level. However, by day 14 the number of colonies increased drastically as compared to the control. Thymocyte supplementation normalized colony formation at any time of observation. Similar pattern is noted in post-radiation repopulation of spleen and bone marrow of mice injected bone marrow pretreated with ABS with or without thymocytes. It is assumed that ABS inactivates bone marrow cells participating in the regulation of CFUs proliferation.     

Secretion of an aluminum chelator, 2-isopropylmalic acid, by the budding yeast, Saccharomyces cerevisiae

An aluminum(III)-binding substance (ABS), that solubilizes Al(III) at neutral pH, was found to be secreted by Saccharomyces cerevisiae. A combination of anion exchange chromatography and preparative high performance liquid chromatography using an octadecylsilane (ODS) column separated ABS from the medium. The structure determination of ABS was performed using 1H and 13C NMR spectroscopy and heteronuclear multiple-bond correlation (HMBC) spectroscopy, and ABS was identified to be 2-isopropylmalic acid (2-iPMA). The structure was further confirmed using high resolution electrospray ionization mass spectrometry. Solubilization of otherwise sparingly soluble Al(III) by 2-iPMA at neutral pH indicated the binding of the compound with Al(III). This is supported by 27Al NMR spectrometry for a solution containing 10 mM Al(III) and 20 mM 2-iPMA at pH 6.6, where four Al(III) species were evident. Although the function of this compound is unclear, it might play a key role in Al detoxification.     

The fetal phenotype of the 18p-syndrome. Report of a male fetus at twenty-one weeks

Morphological and cytogenetic findings in a male fetus at 21 weeks gestation after prenatally detected monosomy 18p are reported. The fetus displayed dysmorphic features resembling the 18p-syndrome, such as decreased head circumference, slightly receding forehead, hypertelorism, epicanthus, horizontal palpebral fissures, depressed nasal bridge, long philtrum, carp mouth, irregular crenated maxillar alveolar ridge, retrognathia, lowset dysplastic ears with posterior rotation, edema of neck, hands and feet respectively, fingers with drop-shaped tips, short first toes with dysplastic nails, hypoplastic male external genitalia. After termination of the pregnancy, biopsies from different fetal organs as well as from the placenta were taken and set up for long term cell cultures. The metaphases of fetal organs all showed the karyotype 46,XY,18p-. A fetal blood culture failed to grow. Unexpectedly, the metaphases of the placenta showed the mosaic karyotype 46,XY/46,XY,18p-/46,XY,18p+.     

Identification of human peripheral T cell antigens.

A new type of differentiation antigens on human T cells was demonstrated by using a heterologous anti-human T cell serum (ATS). This type of antigen, referred to as human peripheral T cell antigen (HPTA), was found on peripheral T cells and medullary thymocytes, but not on cortical thymocytes and B cells. The percentage of ATS-reactive lymphocytes in human peripheral lymphoid organs was correlated with that of cells rosetting with sheep erythrocytes, but contrasted with the number of B cells defined by the presence of a complement (C) receptor or by rabbit anti-human B cell serum (ABS). ATS also reacted with T cells purified by nylon fiber column filtration but ABS did not. Chronic lymphocytic leukemia cells rosetted with either sheep erythrocytes or erythrocyte-antibody-complement complexes were lysed by ATS and ABS, respectively. Mitogenic responses of blood lymphocytes to phytohemagglutinin (PHA) and concanavalin-A (Con A) were abrogated by treating them with ATS and C, whereas ABS suppressed only their response to Con A. Although numerous thymus cells rosetted with SRBC, only 14% were reactive with ATS. Quantitative absorption studies demonstrated that HPTA content of the thymus cells was much lower than that of lymph node cells. Anatomical localization of ATS-reactive lymphocytes in human lymphoid organs studied by immunofluorescence indicated that they were present in the thymus-dependent paracortical areas of lymph node and in the medullary region of thymus. ABS, on the other hand, did not stain thymocytes but reacted selectively with the cells located in the lymphoid follicles of lymph node. These data, together with that from cell suspension studies, confirmed that HPTA were shared between medullary thymocytes and peripheral T cells.     

Observations on the fetus and placenta of a proboscis monkey (Nasalis Iarvatus)

The villous bidiscoid placenta in utero of a proboscis monkey with a fetus of 9.0 cm crown rump length was examined. The immature male fetus weighed 56 g and had a 5.4 cm head circumference. Of particular interest was the stomach, consisting of three parts, large fundus, a narrow tubular portion, and a small pyloric region. The fetal adrenal glands were remarkable in that they were larger than the kidneys. Thus, the adrenal/kidney weight ratio approximated 1.18. The marginally inserted umbilical cord measured 15 cm. Owing to the superficial implantation in the proboscis monkey, penetration of syncytiotrophoblast and a fibrin layer were not observed in the basal plate of the placenta; cytotrophoblastic cell columns attached to the basal plate, consisting of decidua basalis without invasion of trophoblastic giant cells. Intraarterial trophoblastic migration was found in the uterus.     

Adjustment of the fetal head and adult pelvis in modern humans

In nonhuman anthropoids, the anteroposterior (AP) diameters of the fetus are greater than the transverse (TR) diameters and the AP diameters of the pelvic planes are greater than the TR diameters: during labor, therefore, the fetus moves through the birth canal without changing position or orientation. In modern humans, the fetal head at term is encephalized and the fetal chest is flattened. The maternal pelvic inlet is flattened in an AP direction, the sacral promontory and the ischial spines are prominent. As a result, AP<TR at the inlet, but AP>TR at the midpelvis and outlet. In addition, the birth canal presents a marked sacral curvature in the AP direction. The human fetus successfully negotiates the birth canal because the three crucial fetal adaptations: (1) spheroidicity of the presenting part of the fetal head, which allows it to “roll” in the pelvis; (2) mobility of the head and chest in all directions; and (3) a capacity for cranial molding, which adapts fetal head dimensions to pelvic dimensions. The result is that the human fetal head and chest can perform multiple rotational movements in order to always present the greatest fetal diameters to the greatest pelvic diameters. Monkeys show a limited degree of encephalization and suffer from narrow TR pelvic diameters without any possibility of fetal adaptations as shown by humans. Apes also show some encephalization but, because of wider TR diameters in the pelvis, they achieve an easy delivery with no need of fetal adaptations.     

Body measurements correlation and x-ray imaging of three Hystrix javanica fetuses in different fetal development stages

Hystrix javanica is endemic species in Indonesia. Study about fetal development of Hystrix javanica are very rare because of sample limitation. This study was carried out to describe the morphometrics and x-ray analysis of three fetuses in different stage to give basic information about fetal development of Hystrix javanica. Three fetus samples fixed in Bouin’s solution was used in this study. Observation was carried out to identify the characteristic of three fetus samples. This included the pattern of hair, body measurements, body volume, and body weight. X-ray analysis was carried out to know the ossification process in the fetal development. Statistical analysis was carried out using Microsoft 365® Excel program software. Three fetus samples had different specific hair pattern, that was hairless, smooth hairs, and smooth hairs with dense-non dense pattern. Body volume of 1^st, 2^nd, and 3^rd fetus were 23mL, 90mL, and 170mL, respectively. Body weight of 1^st, 2^nd, and 3^rd fetus were 19.5g, 79.22g, and 153.18g, respectively. Pearson’s correlation analysis shown strong relationship between total body length, front body length, back body length, horizontal body diameter, vertical body diameter, head length, and head diameter against body volume and body weight of three fetuses. Significant positive correlation was shown between horizontal body diameter, vertical body diameter, and head diameter against body volume and body length with P value < 0.05. Faint radiopaque images showed in the 2^nd fetus sample and strong radiopaque images showed in the 3^rd fetus sample. Radiopaque images were identified in the teeth, cranium, vertebrae, and extremities bones. In this study we concluded that there was a specific hair pattern in different fetal stage. All body measurements have positive correlation with body volume and body weight and x-ray analysis shown that the ossification of the bone was started to happen while the smooth hair was growth.     

Further evidence for a biological role of anti-estrogen-binding sites in mediating the growth inhibitory action of diphenylmethane derivatives

Several diphenylmethane derivatives have been synthesized with variable affinities for Anti-estrogen Binding Sites (ABS) but not for the estrogen receptor. Using these molecules as probes it is shown that their binding affinities for ABS correlate with their abilities to inhibit the growth of MCF-7 human breast cancer cells. In contrast they have no influence on the proliferation of tamoxifen-resistant variant cells (RTx6) in which ABS are undetectable. These data support the conclusion that ABS has a functional role in the anti-proliferative effect of triphenylethylene anti-estrogens and structurally related compounds.     

闭锁繁育乌骨鸡群体的RAPD指纹分析

随机抽样测定了一个长期闭锁繁育和一个开放繁育雪峰乌骨鸡群体的早期生长和成活率,并藉ＲＡＰＤ分析比较了２个乌骨鸡群体的遗传学特征,结果表明：闭锁繁育群体ＲＡＰＤ标记的平均共带系数和平均带纹频率显著高于开放群体．标记效应估计结果表明随机引物Ｓ１０２扩增的ｂ４（１８２０ｂｐ）和ｂ５（１９３０ｂｐ）片段和Ｓ１０５扩增的ｂ９（１８４９ｂｐ）和ｂ１０（４１０４ｂｐ）片段作为遗传标记对于体重（尤其是３５日龄和７０日龄体重）具有很高的负向相关效应,因此,该４个ＲＡＰＤ标记可以作为近交衰退候选指示标记加以深入研究,最终结论有待于在大样本、多群体、多世代资料中验证．