Recycling coffee grounds and tea leaf wastes to improve the yield and mineral content of grains of paddy rice.

BACKGROUND: Coffee grounds and tea leaf wastes exhibit strong affinity for metals such as Fe and Zn. The objective of this experiment was to evaluate the effect of top-dressing application of Fe- and Zn-enriched coffee grounds and tea leaf wastes at the panicle initiation stage on the mineral content of rice grains and the yield of paddy rice. RESULTS: The Fe and Zn contents of brown rice grains increased significantly on application of both coffee and tea waste materials. The concentration of Mn was increased by top-dressing application of coffee waste material only. For Cu, no significant (P < 0.05) differences were found between the control and ferrous sulfate/zinc sulfate treatment. The application of coffee and tea waste materials led to a significant (P < 0.05) increase in the number of grains per panicle, which was reflected in increases in the total number of grains per hill and in grain yield. CONCLUSION: The top-dressing application of these materials is an excellent method to recycle coffee grounds and tea wastes from coffee shops. Use of these novel materials would not only reduce the waste going to landfill but would also benefit the mineral nutrition of rice consumers at low cost by increasing Fe and Zn levels of rice grains as well as grain yield.

Vitellogenesis in both sexes of gonochoristic mud shrimp, Upogebia major (Crustacea): analyses of vitellogenin gene expression and vitellogenin processing.

The mud shrimp, Upogebia major is a gonochoristic species with distinct sexual dimorphism; however, the male has the "ovarian part of testis" in the gonad and mature-looking eggs appear in a similar reproductive cycle to the female. Vitellogenesis of U. major was investigated focusing on the characterization of vitellogenin (Vg) gene expression and Vg processing. Vg cDNA cloned by PCR-based methods was 7,799 bp-long, encoding 2,568 amino acids in a single open reading frame. The deduced amino acid sequence shared common characteristics conserved in other shrimp Vgs. The Vg gene was expressed in the hepatopancreas of females and males, the ovary, and the ovarian part of testis. Vitellins (Vns) were detected in the gonads of both females and males as three prominent polypeptides with apparent molecular masses of 82 kDa, 100 kDa, and 115 kDa. N-terminal amino acid sequences determined for the three polypeptides were present in the deduced amino acid sequence, demonstrating that they derived from a single long Vg polypeptide. Immunoblot analysis using polyclonal antibodies raised against two Vns (82 kDa and 100 kDa) confirmed Vg processing in the hepatopancreas, in the hemolymph and possibly in the oocytes, similarly in both sexes.

Structural and expression analyses of two vitellogenin genes in the carp, Cyprinus carpio.

We cloned and sequenced two vitellogenin (vg) cDNAs of the carp, Cyprinus carpio, using a cDNA library constructed from estradiol-17 beta (E2)-treated livers. One was a novel, longer 5000 bp-long cDNA termed vg-B2 encoding 1624 amino acids in a single open reading frame. The other was a shorter cDNA (vg-B1), identical to that registered previously as carp vg cDNA in the international nucleotide sequence database. The deduced amino acid sequences of these two molecules were well-aligned with known vertebrate Vgs sharing common characteristics such as N-terminal lipovitellin I (LVI), phosvitin (PV) and C-terminal lipovitellin II (LVII). The novel Vg-B2 bore a highly conserved GL/ICG motif within the LVII region, in contrast to the shorter Vg-B1 that has a truncated C-terminal and lacks the beta-component within the LVII region including the GL/ICG motif. Both vg-B2 and vg-B1 genes were expressed in the livers of females and E2-injected males. Western blot analysis using anti-Vg and anti-vitellin (Vn) antisera demonstrated that both Vg-B2 and Vg-B1 were detected as polypeptides with an estimated molecular mass of 180 kDa and 160 kDa, respectively, in the blood of females and E2-injected males. The results suggest the potential utilization of these genes as sensitive xenoestrogenic markers.

Structure, formation, mechanical properties, and disposal of the embryo attachment system of an estuarine crab, Sesarma haematocheir.

In most decapod crustaceans, fertilized eggs extruded from the gonopore attach to ovigerous hairs within the incubation chamber of the female. The attachment is effected by an "embryo attachment system." The three continuous components of this system are the egg envelope, the funiculus, and the investment coat, which wraps around an ovigerous hair. Transmission electron microscopy (TEM) revealed that the embryo of Sesarma haematocheir is enfolded by three distinct envelopes (E1, E2, and E3), whereas the embryo attachment system is composed of only the outermost, single envelope (E1) with two sublayers (E1a and E1b). This envelope (E1) originates from the outer layer of the vitelline membrane (envelope of the ovum) with two sublayers (E1a' and E1b'). The sequence and timing of events in the formation of the embryo attachment system was determined on the basis of observations of female behavior, ultrastructure, and mechanical properties of the membranes. The egg envelope (E1a' + E1b') is not adhesive immediately after extrusion from the gonopore; but 5 min after egg-laying, it becomes adhesive-a change associated with "fusion" of the two sublayers (E1)-and attaches the eggs to the ovigerous hairs from 5 to 30 min after egg-laying. The layer E1a' always binds to an ovigerous hair at specific, electron-dense attachment sites that are distributed longitudinally on the surface of each hair. Plasticity of the egg envelope changes, and the female kneads her eggs by the movement of ovigerous setae; this movement forms the investment coat on the ovigerous hair (10-40 min after egg-laying). Thirty minutes after egg-laying, the egg envelope again divides into two sublayers (E1a and E1b), and the adhesiveness rapidly decreases. The plasticity of the envelope remains, and the funiculus is formed, accompanied by kneading of the eggs (40-90 min after egg-laying). The embryos hatch one month after incubation, and the attachment systems all slip off their ovigerous hairs by the actions of the ovigerous-hair slipping substance (OHSS). This substance appears to act specifically at the attachment sites on the hair, lysing the bond with layer E1a, and thereby disposing of the embryonic attachment system and preparing the hairs for the next clutch of embryos.

Hatching of an estuarine crab, Sesarma haematocheir: from disappearance of the inner (E3) layer to rupture of the egg case.

Hatching of decapod crustaceans is characterized by the sudden rupture of the egg case. This study focused on the following two issues regarding the hatching mechanism of the estuarine terrestrial crab Sesarma haematocheir: (1) dissolution of the egg case, and (2) the site where the egg case breaks. The egg case comprises three layers: the outer two (E1 and E2) layers and the inner (E3) thin layer (0.2 microm in thickness). The outer layers showed no morphological changes upon hatching, but the inner layer (E3) was markedly digested. The digestion of this layer would enable the embryo to absorb ambient water via reverse peristalsis of the intestine, resulting in an increase of the volume. The egg case always ruptured perpendicular to the longitudinal axis of the embryo. In addition, breakage of the egg case occurred at the dorsal thorax of the embryo. The three major organs positioned at this area were (1) a sharp projection (dorsal spine), (2) an assemblage of muscles, and (3) a pair of secretory glands, each of which was about 30 microm in diameter. The dorsal projection is soft before hatching, and it is clear that the egg case does not break with the posterior expansion of this projection. The rupture instead appears to be caused by the expansion of the muscles arranged perpendicular to the body axis. In addition, some (unknown) factor might weaken the egg case just before hatching. The secretory glands may be a kind of rosette gland, but the role that this gland plays at hatching is not known. As a duct comes out from the center and enters the dorsal projection, some active substance may be released at the tip of this projection. However, immunochemical studies are not consistent with this substance being an ovigerous hair stripping substance (OHSS).

Incidence of tracheal stenosis and other late complications after percutaneous tracheostomy.

OBJECTIVE: To determine the incidence of tracheal stenosis, voice and breathing changes, and stomal complications after percutaneous dilatational tracheostomy (PDT). METHODS: From December 1992 through June 1999, 420 critically ill patients underwent 422 PDTs. There were 340 (81%) long-term survivors, 100 (29%) of whom were interviewed and offered further evaluation by fiberoptic laryngotracheoscopy (FOL) and tracheal computed tomography (CT). Tracheal stenosis was defined as more than 10% tracheal narrowing on transaxial sections or coronal and sagittal reconstruction views. Forty-eight patients agreed to CT evaluation; 38 patients also underwent FOL. CT and FOL evaluations occurred at 30 +/- 25 (mean +/- standard deviation) months after PDT. RESULTS: Twenty-seven (27%) patients reported voice changes and 2 (2%) reported persistent severe hoarseness. Vocal cord abnormalities occurred in 4/38 (11%) patients, laryngeal granuloma in 1 (3%) patient, focal tracheal mucosal erythema in 2 (5%) patients, and severe tracheomalacia/stenosis in 1 (2.6%) patient. CT identified mild (11-25%) stenosis in 10 (21%) asymptomatic patients, moderate (26-50%) stenosis in 4 (8.3%) patients, 2 who were symptomatic, and severe (>50%) stenosis in 1 (2%) symptomatic patient. Ten patients (10%) reported persistent respiratory problems after tracheal decannulation, but only four agreed to be studied. Two patients had moderate stenosis, and one had severe stenosis. One patient's CT scan was normal. No long-term stomal complications were identified or reported. CONCLUSIONS: Subjective voice changes and tracheal abnormalities are common after endotracheal intubation followed by PDT. Long-term follow-up of critically ill patients identified a 31% rate of more than 10% tracheal stenosis after PDT. Symptomatic stenosis manifested by subjective respiratory symptoms after decannulation was found in 3 of 48 (6%) patients.

Ovigerous-hair stripping substance (OHSS) in an estuarine crab: purification, preliminary characterization, and appearance of the activity in the developing embryos.

Ovigerous-hair stripping substance (OHSS) is an active factor in crab hatch water (i.e., filtered medium into which zoea larvae have been released). This factor participates in stripping off the egg attachment structures (i.e., egg case, funiculus, and the coat investing ovigerous hairs) that remain attached to the female's ovigerous hairs after larval release. Thus this activity prepares the hairs for the next clutch of embryos. OHSS activity of an estuarine crab, Sesarma haematocheir, eluted as a single peak on molecular-sieve chromatography, but this peak still showed two protein bands at 32 kDa and 30 kDa on SDS-PAGE. The two protein bands stained with a polyclonal antiserum raised to the active fractions from molecular-sieve chromatography. Moreover, antibodies purified from this polyclonal OHSS antiserum also recognized both the 32-kDa and 30-kDa bands. OHSS immunoreactivity and biological activity were associated with the attachment structures that remained connected to the ovigerous hairs after hatching. In developing embryos, both protein bands could be stained immunochemically at least 10 days before hatching. But OHSS biological activity appeared only 3 days before hatching. The immunoreactive protein bands were not observed in the zoea, but OHSS bioreactivity was present, though greatly reduced. The 32-kDa protein, at least, is probably an active OHSS, and the 30-kDa protein band may also be OHSS-related. The OHSS appears to be produced and stored by the developing embryo. Upon hatching, most of the material may be trapped by the remnant structures, and the remainder is released into the ambient water.

Clinical evaluation of hydroxyapatite-coated titanium plasma-sprayed and titanium plasma-sprayed cylinder dental implants: a preliminary report.

OBJECTIVE: The purpose of this article was to present the preliminary results of a prospective clinical trial comparing titanium plasma-sprayed versus hydroxyapatite-coated titanium plasma-sprayed cylinder (press fit) implants in different regions of the mouth. STUDY DESIGN: Sixty-five subjects met the inclusion requirements. Surgery was done in two phases by four experienced surgeons. Implant placement and abutment connection were separated by 3 to 4 months in the mandible, 6 to 7 months in the maxilla. Patients were assigned to either titanium plasma-sprayed or hydroxyapatite-coated implants on the day of surgery. Implant placement was not stratified for the region of the jaws. Outcome assessment was failure (loss) of an implant before or within 3 months of second phase surgery. RESULTS: Three hundred fifty-two implants equally distributed between titanium plasma-sprayed and hydroxyapatite-coated titanium plasma-sprayed implants were placed in four different sites; anterior maxilla, posterior maxilla, anterior mandible, and posterior mandible. There were a total of 15 failures (4.26%). Overall, titanium plasma-sprayed implants showed a higher but not significant failure rate compared with hydroxyapatite-coated implants (p = 0.06). Although not statistically significant, we believe that a smoking history played an important role in the failure of implants. CONCLUSION: This study suggests that an hydroxyapatite-coating of an implant allows superior initial integration when compared with a titanium plasma-sprayed surface.

Two Kinds of Active Factor in Crab Hatch Water: Ovigerous-Hair Stripping Substance (OHSS) and a Proteinase.

The embryos of intertidal and estuarine crabs are clustered on the ovigerous seta of the female, where they are ventilated for 2-4 weeks by the female's abdomen. When the embryonic development is complete, hatching occurs and zoea larvae are released into the water. This study indicates that the crab hatch water (i.e., the filtered medium into which zoeas were released) contains at least two kinds of active substance: OHSS (ovigerous-hair stripping substance) and a proteolytic enzyme. Both factors were separated by gel filtration. Powdered fragments of egg capsule were digested by proteinase, suggesting that this enzyme actually acts on the egg capsule. But this activity was at a very low level compared with casein digestion. The proteinase might be digesting the thin, sticky layer enclosing the embryo and would not act on the thick, tough layer constituting the main component of the egg capsule. Therefore, a proteolysis of such low activity could not be expected to cause the egg capsule to rupture.

Bioassay and Preliminary Characterization of Ovigerous-Hair Stripping Substance (OHSS) in Hatch Water of Crab Larvae.

Hatch water (the filtrated medium into which zoea larvae have been released) of the estuarine terrestrial crab Sesarma haematocheir (akate-gani) contains a substance that causes premature detachment of embryos from ovigerous females. Detachment occurs when the ovigerous hairs along the female's ovigerous setae slip out of the investment coat that binds them to the embryos through stalks, or funiculi. The active factor, which I call ovigeroushair stripping substance (OHSS), is released outside of the egg capsule at the time of hatching, and is not secreted by the female. This study describes the results of a quantitative assay for measuring the activity of OHSS. Activity is measured as the percentage of hairs on a seta that can be induced to slip out of the coat without damage. Experiments with an extract of crushed embryos indicated that OHSS is present up to 2 days before hatching. Its activity was destroyed by heat and trypsin, suggesting that it is a protein. Its molecular size was estimated by gel filtration to be 15-20 kDa in S. haematocheir and 30 kDa in S. pictum. Reciprocal tests among different species indicated that OHSS occurs widely in intertidal and estuarine crabs.

The Tidal Rhythm of Emergence, and the Seasonal Variation of This Synchrony, in an Intertidal Midge.

The emergence of an intertial midge was investigated at a site on the coast of the Inland Sea of Japan. The population emerging at this site was drawn from a single species of the genus Clunio, probably C. tsushimensis. Emergence was not synchronized with the day-night cycle, but with the tidal cycle. Moreover, the pattern of synchrony changed with season. A bimodal phase appeared in midwinter; but the pattern of synchrony shifted gradually, during January and February, from morning low tides to afternoon low tides, and a unimodal phase appeared in March. This pattern--i.e., synchrony with afternoon low tides--lasted until early October. In mid-October, the synchrony shifted to the morning low tides. Only a brief bimodal phase appeared in autumn. The phase modality was clearly correlated with the height of tides; i.e., when the low waters in a day were very different in height, emergence was synchronized only with the lower one (April to December). During January and February, the higher low tide, as well as the lower low tide, recedes considerably. The exposure of the larval habitat at the higher low tide may stimulate emergence, resulting in bimodal phases in midwinter. But the unimodal pattern in March cannot be accounted for by a simple synchrony with lower low tide, or with exposure of the larval habitat to the air; the day-night cycle not only would be one of the zeitgebers of the tidal rhythm in every season, but also must participate in the expression of the unimodal phase in spring. Furthermore, the number of midges that emerged each day fluctuated with a semilunar cycle with the season. The phase of this rhythm would be shifted by water temperature.

A Substance Inducing the Loss of Premature Embryos From Ovigerous Crabs.

The embryos of an estuarine terrestrial crab (akate-gani; Sesarma haematocheir) are attached by a funiculus to ovigerous hairs on the maternal pleopods, and are ventilated by the female until hatching occurs. When females were kept in a small quantity of diluted seawater (about 10%), hatching and larval release occurred in all cases. In contrast, when the medium was hatch water (i.e., the filtered medium into which larvae had been released), most ovigerous females liberated their embryos prematurely, but hatching did not occur. The egg masses (cluster of embryos) carried by these females were not released as usual, but were gradually extruded from the brooding chamber, and within a few days all had dropped to the bottom of the beaker. No morphological changes were found on the outer egg membrane, the funiculus, or the coat investing the ovigerous hairs of females kept in hatch water. But the ovigerous hairs did slip easily out of the coat, and this caused the extrusion of the egg masses. The active factor--called incubation disrupting substance--was stable with freezing, but heat-labile. In normal females (i.e., those not treated with hatch water), broken egg cases and funiculi remain for a time after hatching with the coat on the ovigerous hairs, but they are gone by the morning after hatching. So the secretion of this incubation-disrupting substance may participate in cleaning the ovigerous hairs of old investing coats and funiculi after larval release, thus preparing for the attachment of the next clutch of embryos. In addition, this substance may play a role in hatching.

Control of Hatching in an Estuarine Terrestrial Crab. II. Exchange of a Cluster of Embryos Between Two Females.

The eggs of an estuarine terrestrial crab, Sesarma haematocheir (akate-gani), are incubated by the female for about one month. In estuarine crabs larval hatching is synchronized with the nocturnal high tide. To investigate whether the female or the embryo controls the actual timing of the hatching, one cluster of embryos was detached from each of two ovigerous females and reciprocally transplanted. Hatching of the transplanted embryos was divided into the following three patterns according to the number of nights until either (or both) of the females released their larvae. In Pattern I, the transplanted clusters both hatched on the same night that the donor females released their larvae. In Pattern II, the hatching of one of the transplanted clusters was not controlled by the host female, whereas hatching of the other transplanted cluster was obviously induced. Finally, in Pattern III, not only the induction of hatching, but also the time of hatching, was controlled by the female. Hatching profiles of transplanted embryos transferred to aerated conditions indicated that hatching requires three nights, and that each embryo also has an endogenous rhythm for hatching. The female seems to play two roles in hatching: i.e., initiation of the hatching process, and enhancement of hatching synchrony in each embryo. A plausible hypothesis explaining the mechanism of induction and the synchronization of hatching is presented.

Control of Hatching in an Estuarine Terrestrial Crab I. Hatching of Embryos Detached From the Female and Emergence of Mature Larvae.

Embryos of the terrestrial crab Sesarma haematocheir hatch simultaneously just prior to their release into water. Larval hatching occurs synchronously when the embryos are attached to a female, and the role of the female in this synchrony has been investigated. Clusters of embryos (200-2000 berries in each cluster) were detached from ovigerous females, and their hatching was compared with that of embryos attached to the females. Of the detached embryos in a cluster, either all hatched, or none hatched. A remarkable feature was that the success of hatching of these detached eggs depended upon the time of hatching of the eggs still attached to the female. Clusters of embryos that were detached from the female within 48-49.5 h of the projected time of larval release all hatched successfully, and swimming zoeas appeared. But embryos that had been detached from the female for longer periods did not hatch at all, though they were obviously alive. These results suggest a hatching process different from the embryonic development process. The female may trigger this process. In addition, detached eggs hatched later than eggs attached to the female, and their hatching was less synchronized. These observations suggest that the female not only initiates hatching, but also enhances the synchrony of hatching.

Phase Shift of a Tidal Rhythm by Light-Dark Cycles in the Semi-Terrestrial Crab Sesarma pictum.

The larval release activity of the semi-terrestrial crab Sesarma pictum was monitored for three-week periods under laboratory conditions of constant and cyclic light. Under conditions of constant dim light, the rhythm for the first ten days was unimodal (larval release just after the nocturnal high tide) and then became bimodal (no apparent synchrony with the tides or with other members of the population) for the remainder of the experimental period. On the other hand, in photoperiods similar to those in the field, the rhythm was maintained; the phase was bimodal and the timing of larval release was delayed 1-2 h from the predicted times of high water in the habitat. When the photoperiod was advanced or delayed, the tidal rhythm was phase-shifted accordingly. The photoperiod does entrain the release rhythm to bimodal tidal cycle. So the phase-shift of a tidal rhythm by 24-h LD cycles is a very difficult phenomenon to explain.

[Synergistic activity of isepamicin and beta-lactam antibiotics against Pseudomonas aeruginosa in vitro and in vivo].

Synergistic activities of isepamicin (ISP) and a beta-lactam antibiotic such as piperacillin (PIPC) or cefotaxime (CTX) against Pseudomonas aeruginosa were demonstrated in vitro and in vivo. In vitro synergistic activity was observed when ISP was used together PIPC or CTX. The synergy observed in vitro was reproduced in vivo against experimental mouse infections, and a ISP-PIPC or a ISP-CTX combination showed significantly greater protective effects than individual antibiotics by themselves.