Quantum cascade lasers monolithically integrated on germanium.

Silicon (Si) photonics can have a major impact on the development of mid-IR photonics by leveraging on the reliable and high-volume fabrication technologies already developed for microelectronic integrated circuits. Germanium (Ge), already used in Si photonics, is a prime candidate to extend the operating wavelength of Group IV-based photonic integrated circuits beyond 8 µm, and potentially up to 15 µm. High performance quantum cascade lasers (QCLs) and interband cascade lasers grown on Si have been demonstrated, whereas no QCLs monolithically integrated on Ge have been reported yet. In this work, we present InAs-based QCLs directly grown on Ge by molecular beam epitaxy. The lasers emitting near 14 µm exhibited threshold current densities as low as 0.8-0.85 kA/cm2 at room temperature.

Interband cascade Lasers with AlGaAsSb cladding layers emitting at 3.3†µm.

We investigate the impact of the growth conditions of AlGaAsSb cladding layers on the properties of interband cascade lasers (ICLs). For an optimized structure emitting at 3.3 µm, we achieve an internal quantum efficiency of 65% per stage in good agreement with conventional ICL using InAs/AlSb superlattice cladding layers, in spite of internal losses of 15 cm-1 due to higher optical losses in the n-type AlGaAsSb alloys. Finally, we report a narrow ridge ICL emitting at 3.33 µm operating in continuous wave up to 80°C that produces 1 mW/uncoated facet at 80 °C, 10 mW at 40 °C and more than 12 mW at 20°C.

High temperature operation of far infrared (λ ≈20 µm) InAs/AlSb quantum cascade lasers with dielectric waveguide.

We demonstrate the high temperature operation, up to 80°C, of quantum cascade lasers emitting at a wavelength of 20 µm. The lasers are based on the InAs/AlSb materials and take benefit of a low loss plasmon-enhanced dielectric waveguide. The waveguide consists of doped InAs cladding layers and low-doped InAs spacers. For 2.9-mm-long devices, the threshold current density is 4.3 kA/cm2 and the measured peak output power is 7 mW at room temperature. The cavity length dependence of the threshold currents also indicates that very large optical gain is achieved and effectively overcome the strong free carrier absorption.

Quantum cutting in Li (770 nm) and Yb (1000 nm) co-dopant emission bands by energy transfer from the ZnO nano-crystalline host.

Li-Yb co-doped nano-crystalline ZnO has been synthesized by a method of thermal growth from the salt mixtures. X-ray diffraction, transmission electron microscopy, atomic absorption spectroscopy and optical spectroscopy confirm the doping and indicate that the dopants may form Li-Li and Yb(3+)-Li based nanoclusters. When pumped into the conduction and exciton absorption bands of ZnO between 250 to 425 nm, broad emission bands of about 100 nm half-height-width are excited around 770 and 1000 nm, due to Li and Yb dopants, respectively. These emission bands are activated by energy transfer from the ZnO host mostly by quantum cutting processes, which generate pairs of quanta in Li (770 nm) and Yb (1000 nm) emission bands, respectively, out of one quantum absorbed by the ZnO host. These quantum cutting phenomena have great potential for application in the down-conversion layers coupled to the Si solar cells.

Stimulated Raman scattering of light in suspension of diamond microparticles in ethanol and in water.

The spectra of stimulated Raman scattering of light in ethanol and in water suspensions containing diamond microparticles with sizes 0.2-0.3 µm were investigated. An excitation radiation source was a pulsed ruby laser with a generation wavelength λ0 = 694.3 nm, a pulse duration τp ≈ 20 ns, a maximum beam energy of Emax = 0.6 J, a spectral width Δν = 0.015 cm-1, and a beam divergence 3.5·10-4 rad. For the first time, the observation of stimulated Raman scattering of light at a boson peak in suspension of diamonds microcrystals with close sizes (0.2-0.3 µm) in a liquid is reported. The corresponding spectra were recorded using a Fabry-Perot interferometer. In this case, the frequency shift of the stimulated Stokes Raman scattering depended on the size of the diamond microparticles introduced into the liquid and amounted to ~1 cm-1. In addition, stimulated Raman scattering by a fundamental optical mode with a frequency shift ν = 1331 cm-1 was observed. In this case, the Raman spectra were recorded using a small-sized spectrometer with a multi-element receiver, detecting radiation in the range of 200-1000 nm. At a sufficiently high intensity of the exciting radiation, the Stokes and anti-Stokes satellites were simultaneously present in the spectrum of stimulated Raman scattering. The obtained results on stimulated scattering of diamond microparticles in liquids are of interest for estimating the sizes of microcrystals from scattering spectra at a boson peak, as well as for creating a frequency comb of emitters based on stimulated Raman scattering with a large frequency shift.

[Molecular genetic basis of proximal spinal muscular atrophy and experience in its pharmaceutical treatment].

The review considers the original and published data on the molecular genetic basis of proximal spinal muscular atrophy (SMA), the most common monogenic neuromuscular disease. The structures of the SMN1 gene and SMN2 pseudogene, mutations distorting the SMN1 function, the structure and functions of the Smn neurotrophic protein, its role in biogenesis of small nuclear ribonucleoproteins (snRNPs), and the principles and prdblems of molecular diagnosis in SMA are described. Special consideration is given to the current approaches and prospects of gene and cell therapy of SMA, pharmacogenetic methods to correct the SMN2 function, and original results of long-term treatment of SMA patients with valproic acid drugs.

[Lysine dendrimers as vectors for delivering genetic constructs to eukaryotic cells].

Asymmetrical lysine dendrimers are promising as vectors for delivering gene expression constructs into mammalian cells. The condensing, protective, and transfection properties were studied for pentaspherical lysine dendrimer D5 and its analog D5C10, modified with capric acid residues at the outer sphere; in addition, the transfection activity was assayed for complexes DNA-dendrimer-endosomolytic peptide JTS-1. Fatty acid residues incorporated in lysine dendrimers proved to improve their ability to bind DNA, to protect DNA from nuclease degradation, and to ensure its transfer into the nucleus. Peptide JTS-1 introduced in DNA-dendrimer complexes significantly increased their transfection activity. The potentiating effect of JTS-1 was especially high with the DNA-D5C10 complex. An excess of JTS-1 changed the structure of the complexes and reduced their transfection activity. It was assumed that dendrimers D5 and D5C10 are promising vectors for delivering DNA to eukaryotic cells and provide a basis for constructing more refined nonvirus module carriers.

[Optimization of transfection properties of DNA-lysine dendrimer complexes].

We studied the possibility of optimizing the DNA transfection properties of carriers based on lysine dendrimers of the third and the fifth generation, including those containing a chloroacetyl or a lipophilic palmitoyl moiety at C-end. The use of lysosome-destroying antibiotic chloroquine and an amphipathic polycationic nonadecapeptide JTS-1 was found to enhance the DNA transfecting properties of the lysine dendrimers. The triple complex including DNA, a lysine dendrimer of the third generation modified with lipophylic moieties of palmitic acid at its C-end, and JTS-1 was shown to be comparable in its transfecting activity to a complex containing Escort, a commercial cationic liposome carrier.

Trace gas detection with antimonide-based quantum-well diode lasers.

Widely tunable GaInAsSb/AlGaAsSb quantum well (QW) lasers have been grown by molecular beam epitaxy on GaSb substrates. Their emission wavelength, from 2.0 to 2.5 microm, make them suitable for the detection of many gas species in the wavelength range which corresponds to an atmospheric transmission window. Using these devices an experimental setup for open path gas detection has been developed.

[Suppression of nonsense mutations in the Dystrophin gene by a suppressor tRNA gene]. / Ispol'zovanie gena supressornoi tRNK dlia ispravleniia nonsens-mutatsii v gene distrofina.

Nonsense mutations in the dystrophin gene are the cause of Duchenne muscular dystrophy (DMD) in 10-15% of patients. In such an event, one approach to gene therapy for DMD is the use of suppressor tRNAs to overcome the premature termination of translation of the mutant mRNA. We have carried out cotransfection of the HeLa cell culture with constructs containing a suptRNA gene (pcDNA3suptRNA) and a marker LacZ gene (pNTLacZhis) using their polymer VSST-525 complexes. It was found that the number of cells producing beta-galactosidase depends inversely on the dose of the suptRNA gene. A single in vivo injection of the construct providing for expression of the suptRNAochre gene into mdx mouse muscle resulted in the production of dystrophin in 2.5% of fibers. This suggests that suppressor tRNAs are applicable in gene therapy for hereditary diseases caused by nonsense mutations.

[Dendrimers based lysine and their "starburst" polymeric derivatives: prospects of use in compacting DNA and in vitro delivery of genetic constructs]. / Dendrimery na osnove lizina i ikh "zvezdoobraznye" polimernye proizvodnye: vozmozhnost' ispol'zovaniia dlia kompaktizatsii DNK i dostavke ékspressiruiushchikh geneticheskikh konstruktsii in vitro.

We attempted to find some compounds for the effective delivery of gene constructs into cells and obtained two trispherical dendrimers on the basis of lysine, (Lys)8-(alpha, epsilon-Lys)4-(alpha, epsilon-Lys)2-(alpha, epsilon-Lys)-Ala-NH2 (D1) and (Lys)8-(alpha, epsilon-Lys)4-(alpha, epsilon-Lys)2-(alpha, epsilon-Lys)-Ala-[Lys(Plm)]2-Ala-NH2 (D2), as well as the starburst polymeric derivatives of D1, (pVIm)8-D1 and (pLys)n-D1, containing poly(N-vinylimidazole) and polylysine chains bound at a single point to the dendrimer amino groups. The conditions of dendrimer-plasmid DNA complex formation were studied. The intracellular localization of these complexes and the expression of gene constructs delivered with their help were analyzed in transfection experiments on the HeLa cell cultures of human epithelial carcinoma and on C2C12 mouse myoblasts. It was found that the chemical structure of dendrimer D1 and its derivatives significantly affected the structure and properties of complex. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2004, vol. 30, no. 1; see also http://www.maik.ru.

[Current state and prospects for gene therapy of Duchenne muscular dystrophy in the world and in Russia]. / Sobremennoe sostoianie i perspektivy gennoi terapii miodistrofii Diushenna v mire i Rossii.

Failure of drug therapy of Duchenne muscular dystrophy (DMD) stimulated intense search for adequate methods of gene therapy (GT) which would ensure effective delivery of the dystrophin (D) gene, its long-term persistence in transfected cells, and its expression in muscle fibers. The main results of the experimental GT of DMD with the use of viral and nonviral delivery of the D gene into muscles of biological models are discussed. Delivery of a mini-gene of D with a specific muscle promoter using a modified adenoassociated virus is currently the most promising method, which will soon be available for clinical trials. The main results of the studies on the DMD GT in Russia are summarized. The results of experiments on genetic transfection of mdx mice with marker genes and various constructions with the D gene are outlined. The genes are delivered into muscles by means of gene gun, electroporation, viral oligopeptides, liposomes, microspheres, lactoferine, and other nonviral vehicles. It is emphasized that consolidation of funds and efforts of all Russian laboratories dealing with gene and cell therapy of DMD are necessary to complete the experiments and start clinical trials.

[Features of transfection and expression of cDNA for the human dystrophin gene, delivered into muscles of mdx mice using MF-2 synthetic microspheres]. / Osobennosti transfektsii i ékspressii kDNK gena distrofina cheloveka, dostavlennogo v myshtsy myshei mdx s pomoshch'iu sinteticheskikh mikrosfer MF-2.

The number of dystrophin-positive myofibers (DPM), that appeared in different skeletal muscles of mdx mice after a single injection of synthetic microspheres containing the full-length human dystrophin cDNA within the pHSADy expressing vector into femoral quadriceps muscle, was examined on cryostat sections. Injection of 25 micrograms cDNA resulted in the occurrence of 1, 2.4, 5.8 and 4.8% of DPM in the treated muscle in 1, 7, 21, and 60 days after the injection respectively. 7, 21, and 60 days after the treatment, these values comprised 2.1, 4.3 and 1% in the same muscle of the contralateral leg, and 5.5, 8.4, and 1% in the gluteal muscle. Expression of the full-length human dystrophin (427 kDa) in the muscle of the transfected mdx mice was observed. The presence of the transfected construction in skeletal muscles, heart, brain, lungs, and fetuses was demonstrated PCR. Utilization of the FISH technique with biotinilated pHSADy construct as a DNA probe showed that 7 days after the injection, the MF-2 microspheres were present in 70% of myoblast nuclei, in 64% of nuclei of gluteal muscles, and in 62% of the contralateral quadriceps nuclei. 21 days after the treatment, these values were 41, 29, and 45%, respectively. The MF-2 microsphere were detected in the nuclei of the blood, brain, heart, and lung cells, as well as in the placenta and tissues of 18-day-old fetuses. Our results demonstrated the high efficiency of microsphere-mediated transfer of gene constructs into cell nuclei, their long-term intranuclear persistence, and the ability to direct expression for at least 2 months after injection. The MF-2 microspheres attract special interest in respect to the targeted delivery of gene constructs into the nuclei.

[Dependence of the efficacy of transfecting muscle fibers in vivo on electroporation conditions]. / Zafisimost' éffektivnosti transfektsii myshechnykh volokon in vio ot uslovii élektroporatsii.

This study is a survey of in vivo experiments on transfection of laboratory mouse muscle fibers by electroporation using an original device generating electric impulses. Transfection efficiency proved to depend on DNA dose and the number of electric impulses. It can be increased significantly by electroporation at varying pulse burst polarity. At both direct electrode application to muscles and electroporation through the skin, the muscle fiber transfection was more efficient under electroporation conditions much milder than those usually reported. The use of electroporation method for gene therapy of Duchenne muscular dystrophy is discussed.

[Expression of the human dystrophin gene in mdx mouse muscle fibers after transfection using liposomes and synthetic oligopeptides]. / Ekspressiia gena distrofina cheloveka v myshechnykh voloknakh myshei mdx posle transfektsii s pomoshch'iu liposom i sinteticheskikh oligopeptidov.

The number of dysrophin-positive fibers appearing in the femoral quadriceps muscle of mdx mice after injection of the full-length human dystrophin cDNA within the pHSADy plasmid was examined by means of immunohystochemical techniques. Transfection was carried out using lipofectamine (LFA), or synthetic oligopeptide complexes that provided the condensation of plasmid DNA (K8) and its release from endosomes gopeptide complexes that provided the condensation of plasmid DNA (K8) and its release from endosomes (JTS1). The LFA + pHSADy at a dose of 10 micrograms DNA did not affect the number of dystrophin-positive fibers at the site of injection (0.6-0.8%), whereas it caused a statistically significant increase in the number of these fibers in the same muscle of the contralateral leg (up to 2.3%). Injection of the SO + pHSADy complex resulted in the occurrence of dystrophin-positive muscle fibers characterized by a heterogeneous content and the distribution of dystrophin. The greatest number of dystrophin-positive fibers (about 16%) was observed under a ratio of pHSADy to K8 of 1:3 or 1:4. The observed maximal number of dystrophin-positive fibers after a single injection of SO + pHSADy was 3.8%, and it was 17.7% after three injections. These values were statistically significantly higher compared to intact mice (0.6%), the injection of pure plasmid (2.2%), or the intramuscular injection of sucrose (from 0.7 to 1.3%). A relatively high level of transfection (about 5%) was observed after an intracardiac injection of a large dose of the pHSADy (70 micrograms DNA). The perspectives of the targeted delivery of the dystrophin gene into muscles under conditions of parenteral administration are discussed.

[Expression of the human dystrophin gene in mdx mouse skeletal muscles after ballistic transfection]. / Ekspressiia gena distrofina cheloveka v skeletnykh myshtsakh myshei mdx posle ballisticheskoi transfektsii.

"Gene-gun" ballistic transfection (BT) was used to deliver genetic constructs pMLVDy and pHSADy containing full-length cDNA of the dystrophin gene to musculus quadriceps remoris and musculus gluteus of mdx mice, which represent a natural model of Duchenne muscular dystrophy. Clusters of dystrophin-positive muscular fibers (DPMF) were immunocytochemically detected in sites exposed to BT. The average number of DPMF was 2% by the 17th day and 3% by the 60th day after BT with pMLVDy, whereas the number of revertant DPMF was 0.2% in control mice (without BT). When pHSADy was used, the average number of DPMF was 3% 20 days after BT. In this case, dystrophin was uniformly spread though the myoplasm in 3% of cells and produced a slight signal in separate regions under the sarcolemma in 10% of muscle fibers. The number of revertant DPMF increased to 0.6% after BT with naked particles and to 2.8% after BT with the marker lacZ gene, in both bombarded and contralateral legs. The number of DPMF in the corresponding muscles of the contralateral leg significantly increased and reached 2.8% by the 60th day after BT with pMLVDy and 6.7% by the 20th day after BT with pHSADy. Human dystrophin gene cDNA was detected in all skeletal muscles, heart, intestine, tongue, and brain by polymerase chain reaction (PCR) three weeks after BT. Immunoblot analysis showed that normal 427-kDa human dystrophin was synthesized in muscles of mdx mice. The results suggest applicability of BT for delivery of dystrophin constructs into muscles.

[The BCL-xL and ACR-1 genes promote differentiation and reduce apoptosis in muscle fibers of mdx mice]. / Geny BCL-xL i ACR-1 sposobstvuiut differentsirovke i umen'shaiut gibel' myshechnykh volokon myshei mdx.

The effects of the human BCL-xL and ACR-1 genes on dystrophin expression in cross-striated muscle fibers (CSMF) and on CSMF viability were studied in mdx mice after ballistic cotransfection with the human dystrophin minigene. In control mice, the proportion of dystrophin-positive (D(+)) and dying CSMF were 2.1 +/- 0.1 and 2.1 +/- 0.3%, respectively. Introduction of the dystrophin minigene (20 micrograms of the pSG5dys plasmid) increased the proportions of D(+) and dying CSMF to 5.6 +/- 1.4% and 4.5 +/- 0.9%, respectively. When pSG5dys was introduced along with the pSFFV-Neo plasmid carrying the BCL-xL gene (10 micrograms of each plasmid per shot), the death of CSMF decreased to 3.7 +/- 1% and the proportion of D(+) CSMF significantly (P < 0.05) increased to 12.2 +/- 2.2%. Contransfection with the dystrophin minigene and the BCL-xL gene at 20 micrograms of each plasmid per shot did not stimulate generation of D(+) CSMF, but did reduce the CSMF death to 1.5 +/- 0.3%. Introduction of pSG5dys along with the pRc-CMV-10.1 plasmid containing the ACR-1 gene (10 micrograms of each plasmid per shot) reduced the proportion of D(+) CSMF to 1.1 +/- 0.5% and significantly reduced the proportion of dying CSMF to 0.9 +/- 0.3% as compared with the proportions observed in intact mice or in mice subjected to transfection with pSG5dys. Introduction of the pSG5dys plasmid substantially reduced the proportion of CSMF with peripheral nuclei, suggesting disturbed CSMF differentiation. After cotransfection with the human-dystrophin minigene, the BCL-xL and ACR-1 genes did not affect the extent of CSMF differentiation as compared with that observed in the case of the dystrophin minigene alone. Thus, ballistic transfection of mdx mice with the human dystrophin gene used along with the BCL-xL or ACR-1 gene was shown to suppress the death of muscle fibers and to expedite dystrophin synthesis and cell differentiation.

[Gene hACR-1 suppresses apoptosis of striated muscle fibres of m. quadriceps femoris after ballistic transfection of mdx mice]. / Gen hACR-1 podavliaet apoptoz poperechnopolosatykh myshechnykh volokon m.quadriceps femoris myshei mdx pri ballisticheskoi transfektsii.

Human minidystrophin gene (pSG5dys plasmid) and hACR-1 gene (pRc-CMV-10.1 plasmid) were cotransfected by means of "gene-gun" to M. quadriceps femoris of mdx mice. Effects of transfection on dystrophin expression and survival of striated muscle fibres (SMF) were studied on the 21st day after shots. In the control mdx dystrophin-positive muscular fibers [D(+)] SMF and destroyed SMF made 2.1 +/- 0.1 and 2.1 +/- 0.3%, respectively. In mice transfected with pSG5dys plasmid (20 mkg of DNA per mouse), the shares of D(+) SMF and dead SMF raised, respectively, up to 5.6 +/- 1.4 and 4.5 +/- 0.9%. Transfection of mice with pRc-CMV-10.1 (DNA dose is 20 mkg per mouse) reduced the levels of apoptosis in SMF and D(+) SMF level to 1.6 +/- 0.6 and 1.1 +/- 0.4%, respectively. Cotransfection by pSG5dys and pRc-CMV-10.1 plasmids (10 and 10 mkg of each plasmids DNA per mouse) reduced the share of D(+) SMF to 1.1 +/- 0.5% and SMF destruction to 0.9 +/- 0.3%. pSG5dys transfection considerably reduced the share of SMF having peripherally located nuclei, thus indicating a decrease in SMF differentiation level after transfection. Cotransfection of ACR-1 gene and a dystrophin minigene did not suppress further cytodifferentiation of mdx muscle fibers. A conclusion is made that ballistic transfection by hACR-1 gene reduces the level of apoptosis in mdx mice SMF without changing the level of SMF differentiation. The cotransfection of mdx mice muscle by hACR-1 and human minidystrophin gene reduces SMF destruction and supports SMF differentiation, too.

[Clinico-anatomic analysis of maternal mortality in the Arkhangelsk region]. / Kliniko-anatomicheskii analiz materinskoi smertnosti v Arkhangel'skoi oblasti.

Clinico-anatomical analysis is presented of 93 autopsies of pregnant women, women in childbirth and puerperas who died in 1985-1999 in the Arkhangelsk region. In this region complications after abortion occur less frequently than in Russia on the whole while the percentage of late gestosis, obstetric hemorrhage and pyoseptic complications were high than mean Russian. In small number of discrepancies between final clinical and anatomo-pathological diagnosis, high percentage of discrepancies was found in the diagnosis of the 3rd category and iatrogenia this being due to difficulties in clinical diagnosis of obstetic pathology and increasing incidence of surgical deliveries.

[Gene therapy of monogenic hereditary diseases. Duchenne myodystrophy]. / Gennaia terapiia monogennykh nasledstvennykh zabolevanii. Miodistrofiia Diushenna.

The paper highlights the new trends in gene therapy research area and clinical trials. It should be noted that the majority of firms involved in development of the scientific approaches to gene therapy are concentrated in the United States. The investments of the given companies on development and research of new genetic constructs also delivery systems make hundred millions dollars. The greatest part (more than 80%) of gene therapy clinical trials projects are also connected with the US research departments; the majority of them is related to tumor therapy. The advantages and drawbacks of the main methods of nucleic acids delivery to the cells are considered; diseases that are attempted to be treated using gene therapy methods are listed. A special attention of the review is devoted to the modern stand in research on cell and Duchenne muscular dystrophy (DMD) gene therapy, also brief description of basic results achieved in the authors laboratory is given. Basic original results of transfection of mdx mice (DMD biological models) with dystrophin cDNA delivered by gene gun, cationic liposomes, synthetic microspheres, viral olygopeptides and lactoferrine are summarized.