Cell specialization within the parenchymatous bundle sheath of barley

Abstract. Structural and physiological aspects of the parenchymatous bundle sheath (PBS) were studied in cultivars of Hordeum distichum L. The PBS of intermediate, lateral and midrib veins consisted of a single layer of cells closely appressed to the mestome sheath. These cells were large, vacuolate and approximately cylindrical in shape, extending parallel to the vein. Mean PBS cell volume was 4 × 10⁻⁵mm³ compared to 1.23 × 10⁻⁵mm³ for mesophyll cells. Transverse sections revealed three cell types within the PBS, cells with small chloroplasts (S-type), cells with large chloroplasts (L-type) and structural cells. The majority of cells were S-type, containing chloroplasts of approximately a third of the volume of mesophyll chloroplasts; they were able to reduce tetranitro blue-tetrazolium and synthesize starch. Structural cells interrupted the phloem and xylem are of the sheath in lateral veins and the midrib, whilst between one and four PBS cells within the phloem are of each vein type contained chloroplasts similar in volume and starch content to those of the mesophyll. Only these L-type cells contained noticeable starch grains at the end of an 8-h dark period, a further 4 h darkness being required for complete mobilization of starch. Starch deposition within S-type and structural cells was detectable after 4 h illumination but was only appreciable in leaves excised from the plant and illuminated for 9–12 h. The role of S-type PBS cells in assimilate transport is discussed in relation to these findings.     

Arabidopsis leaf hydraulic conductance is regulated by xylem sap pH,controlled, in turn,by a P-type H+-ATPase of vascular bundle sheath cells

The leaf vascular bundle sheath cells (BSCs) that tightly envelop the leaf veins, are a selective and dynamic barrier to xylem sap water and solutes radially entering the mesophyll cells. Under normal conditions, xylem sap pH below 6 is presumably important for driving and regulating the transmembranal solute transport. Having discovered recently a differentially high expression of a BSC proton pump, AHA2, we now test the hypothesis that it regulates the xylem sap pH and leaf radial water fluxes. We monitored the xylem sap pH in the veins of detached leaves of wild-type Arabidopsis, AHA mutants and aha2 mutants complemented with AHA2 gene solely in BSCs. We tested an AHA inhibitor (vanadate) and stimulator (fusicoccin), and different pH buffers. We monitored their impact on the xylem sap pH and the leaf hydraulic conductance (K_leaf), and the effect of pH on the water osmotic permeability (P_f) of isolated BSCs protoplasts. We found that AHA2 is necessary for xylem sap acidification, and in turn, for elevating K_leaf. Conversely, AHA2 knockdown, which alkalinized the xylem sap, or, buffering its pH to 7.5, reduced K_leaf, and elevating external pH to 7.5 decreased the BSCs P_f. All these showed a causative link between AHA2 activity in BSCs and leaf radial hydraulic water conductance.     

Predictive Value of Calcium Test for Preoperative Diagnosis of Medullary Thyroid Carcinoma in Patients With Moderately Elevated Basal Calcitonin

ObjectiveMedullary thyroid carcinoma (MTC) can be very aggressive, and early diagnosis is based on routine measurement of serum calcitonin (CT) and RET genetic testing for hereditary forms. Basal serum CT (bCT) concentrations are useful in the early detection of MTC, although it is still unclear whether they can also be used for the differential diagnosis between MTC and C-cell hyperplasia (CCH). Since false-positive results can be obtained with the basal measurement of CT, a provocative test to evaluate stimulated CT (sCT) is often needed. The objective of this study was to investigate the utility of a calcium gluconate test for CT in distinguishing MTC from CCH, a precancerous condition in hereditary forms of MTCs but with unclear significance in sporadic MTCs.MethodsA total of 74 patients underwent the calcium loading test before thyroidectomy, and bCT and sCT levels were compared with histologic results by receiver operating characteristic plot analyses.ResultsA peak CT level of 388.4 pg/mL after stimulation with calcium gluconate was able to significantly distinguish patients with MTC from those with CCH and those without C-cell pathology, with 81.8% sensitivity and 36.5% specificity. A bCT level of 16.1 pg/mL was able to distinguish between these 2 groups of patients with a sensitivity of 90%.ConclusionHigh-dose calcium test is an effective procedure that can be applied for differential diagnosis of MTC and CCH. Reference ranges for calcium sCT levels and CT thresholds in different groups of patients have been identified.     

Lamina specific postnatal development of PKCγ interneurons within the rat medullary dorsal horn

Protein kinase C gamma (PKCγ) interneurons, located in the superficial spinal (SDH) and medullary dorsal horns (MDH), have been shown to play a critical role in cutaneous mechanical hypersensitivity. However, a thorough characterization of their development in the MDH is lacking. Here, it is shown that the number of PKCγ‐ir interneurons changes from postnatal day 3 (P3) to P60 (adult) and such developmental changes differ according to laminae. PKCγ‐ir interneurons are already present at P3‐5 in laminae I, IIo, and III. In lamina III, they then decrease from P11–P15 to P60. Interestingly, PKCγ‐ir interneurons appear only at P6 in lamina IIi, and they conversely increase to reach adult levels at P11–15. Analysis of neurogenesis using bromodeoxyuridine (BrdU) does not detect any PKCγ‐BrdU double‐labeling in lamina IIi. Quantification of the neuronal marker, NeuN, reveals a sharp neuronal decline (∼50%) within all superficial MDH laminae during early development (P3–15), suggesting that developmental changes in PKCγ‐ir interneurons are independent from those of other neurons. Finally, neonatal capsaicin treatment, which produces a permanent loss of most unmyelinated afferent fibers, has no effect on the development of PKCγ‐ir interneurons. Together, the results show that: (i) the expression of PKCγ‐ir interneurons in MDH is developmentally regulated with a critical period at P11‐P15, (ii) PKCγ‐ir interneurons are developmentally heterogeneous, (iii) lamina IIi PKCγ‐ir interneurons appear less vulnerable to cell death, and (iv) postnatal maturation of PKCγ‐ir interneurons is due to neither neurogenesis, nor neuronal migration, and is independent of C‐fiber development. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 77: 102–119, 2017     

Suitable fusion of N-terminal heptad repeats to achieve covalently stabilized potent N-peptide inhibitors of HIV-1 infection

N-terminal heptad repeat (NHR)-derived peptide (N-peptide) fusion inhibitors, which are derived from human immunodeficiency virus (HIV) envelope glycoprotein 41 (gp41), are limited by aggregation and unstable trimer conformation. However, they could function as potent inhibitors of viral infection by forming a coiled-coil structure covalently stabilized by interchain disulfide bonds. We previously synthesized N-peptides with potent anti-HIV-1 activity and high stability by coiled-coil fusion and covalent stabilization. Here, we attempted to study the effects of NHRs of chimeric N-peptides by fusing de novo coiled-coil isopeptide bridge-tethered T21 peptides of different NHR lengths. Peptides (T21N23)₃ and (T21N36)₃ was a more potent HIV-1 fusion inhibitor than (T21N17)₃. The site of isopeptide bond formation was precisely controlled and had little influence on N-peptide properties. The N-peptide (T21N36)₃, which had a similar conformation as the NHR trimer and interacted well with the C34 peptide, may be useful for screening other C-peptides and small-molecule fusion inhibitors, and for studying the interactions between the NHR trimer and C-terminal heptad repeats.     

Systematic and comparative anatomy of Cymbidieae (Orchidaceae)

Cymbideae comprise an assemblage of 28 genera nearly all of which are represented in this study. Their anatomy is relatively homogenous with the exception of Govenia , in which roots lack velamen and pseudobulb vascular bundles lack sclerenchyma, conditions that do not obtain in other genera. Marginal fibre bundles in leaves of Grammatophyllum and Porphyroglottis consist of clusters of thicker-walled, narrower, epidermis-facing fibres as well as thinner-walled, wider, mesophyll-facing fibres. This feature also occurs in some species of Maxillaria . Baculate tilosomes appear in the roots of a majority of genera in Cymbidieae, as they do in species of Maxillaria , confirming DNA analyses showing a close relationship between tribes Cymbidieae and Maxillarieae. Govenia is singled out both on anatomical and molecular grounds as being ill-placed in Cymbidieae. Cladistic analysis produces only a few tentative hypotheses of phylogenetic relationships among the 28 genera, showing that anatomical characters are of limited value in assessing affinities within this tribe. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 139 , 1–27.