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Supplementary Figure from Ontogeny and Vulnerabilities of Drug-Tolerant Persisters in HER2+ Breast Cancer

Resistance to targeted therapies is an important clinical problem in HER2-positive (HER2+) breast cancer. “Drug-tolerant persisters” (DTP), a subpopulation of cancer cells that survive via reversible, nongenetic mechanisms, are implicated in resistance to tyrosine kinase inhibitors (TKI) in other malignancies, but DTPs following HER2 TKI exposure have not been well characterized. We found that HER2 TKIs evoke DTPs with a luminal-like or a mesenchymal-like transcriptome. Lentiviral barcoding/single-cell RNA sequencing reveals that HER2+ breast cancer cells cycle stochastically through a “pre-DTP” state, characterized by a G0-like expression signature and enriched for diapause and/or senescence genes. Trajectory analysis/cell sorting shows that pre-DTPs preferentially yield DTPs upon HER2 TKI exposure. Cells with similar transcriptomes are present in HER2+ breast tumors and are associated with poor TKI response. Finally, biochemical experiments indicate that luminal-like DTPs survive via estrogen receptor–dependent induction of SGK3, leading to rewiring of the PI3K/AKT/mTORC1 pathway to enable AKT-independent mTORC1 activation.Significance:DTPs are implicated in resistance to anticancer therapies, but their ontogeny and vulnerabilities remain unclear. We find that HER2 TKI-DTPs emerge from stochastically arising primed cells (“pre-DTPs”) that engage either of two distinct transcriptional programs upon TKI exposure. Our results provide new insights into DTP ontogeny and potential therapeutic vulnerabilities.This article is highlighted in the In This Issue feature, p. 873

Supplementary Table from Ontogeny and Vulnerabilities of Drug-Tolerant Persisters in HER2+ Breast Cancer

Supplementary Figure 1-23

In lung cancer, enrichment of the lower airway microbiota with oral commensals commonly occurs, and ex vivo models support that some of these bacteria can trigger host transcriptomic signatures associated with carcinogenesis. Here, we show that this lower airway dysbiotic signature was more prevalent in the stage IIIB–IV tumor–node–metastasis lung cancer group and is associated with poor prognosis, as shown by decreased survival among subjects with early-stage disease (I–IIIA) and worse tumor progression as measured by RECIST scores among subjects with stage IIIB–IV disease. In addition, this lower airway microbiota signature was associated with upregulation of the IL17, PI3K, MAPK, and ERK pathways in airway transcriptome, and we identified Veillonella parvula as the most abundant taxon driving this association. In a KP lung cancer model, lower airway dysbiosis with V. parvula led to decreased survival, increased tumor burden, IL17 inflammatory phenotype, and activation of checkpoint inhibitor markers.Significance:Multiple lines of investigation have shown that the gut microbiota affects host immune response to immunotherapy in cancer. Here, we support that the local airway microbiota modulates the host immune tone in lung cancer, affecting tumor progression and prognosis.See related commentary by Zitvogel and Kroemer, p. 224.This article is highlighted in the In This Issue feature, p. 211

Supplementary File Table 1-10

Supplementary Table

Quantum technologies have outgrown mere fundamental research in laboratories over recent years, and will facilitate more and more potentially disruptive applications in a wide range of fields in the future. In foresight, qualification opportunities need to be implemented in order to train qualified specialists, referred to as the future quantum workforce, in various fields. Universities world-wide have launched qualification programmes for engineers focusing on quantum optics and photonics. In many of these programmes, students attend courses on quantum physics contextualized via quantum optics experiments with heralded photons, because: (1) their experimental and physical foundations may be directly leveraged to teaching a number of quantum technology applications, and (2) physics education research has provided empirical evidence, according to which such quantum optics-based approaches are conducive to learning about quantum concepts. While many teachers are confident about the effectiveness of their concepts, there is little empirical evidence due to the lack of content-area-specific research tools. We present a 16-item concept inventory to assess students’ conceptual understanding of quantum optics concepts in the context of experiments with heralded photons adopted from a test instrument published in the literature. We have administered this Quantum Optics Concept Inventory as a post-test to N=216 students after instruction on quantum optics as part of an undergraduate engineering course. We evaluated the instruments’ psychometric quality, both in terms of classical test theory, and using a Rasch scaling approach. The Quantum Optics Concept Inventory enables a reliable measure (α=0.74), and the data gathered show a good fit to the Rasch model. The students’ scores suggest that fundamental quantum effects pose striking learning hurdles to the engineering students. In contrast, most of the students are able to cope with the experimental and technical foundations of quantum optics experiments with heralded photons and their underlying principles, such as the coincidence technique used for the preparation of single-photon states. These findings are in accordance with prior research, and hence, the Quantum Optics Concept Inventory may serve as a fruitful starting point for future empirical research with regard to the education of the future quantum workforce.

Rationale: Workers' exposure to metalworking fluid (MWF) has been associated with respiratory disease.Objectives: As part of a public health investigation of a manufacturing facility, we performed a cross-sectional study using paired environmental and human sampling to evaluate the cross-pollination of microbes between the environment and the host and possible effects on lung pathology present among workers.Methods: Workplace environmental microbiota were evaluated in air and MWF samples. Human microbiota were evaluated in lung tissue samples from workers with respiratory symptoms found to have lymphocytic bronchiolitis and alveolar ductitis with B-cell follicles and emphysema, in lung tissue samples from control subjects, and in skin, nasal, and oral samples from 302 workers from different areas of the facility. In vitro effects of MWF exposure on murine B cells were assessed.Measurements and Main Results: An increased similarity of microbial composition was found between MWF samples and lung tissue samples of case workers compared with control subjects. Among workers in different locations within the facility, those that worked in the machine shop area had skin, nasal, and oral microbiota more closely related to the microbiota present in the MWF samples. Lung samples from four index cases and skin and nasal samples from workers in the machine shop area were enriched with Pseudomonas, the dominant taxa in MWF. Exposure to used MWF stimulated murine B-cell proliferation in vitro, a hallmark cell subtype found in the pathology of index cases.Conclusions: Evaluation of a manufacturing facility with a cluster of workers with respiratory disease supports cross-pollination of microbes from MWF to humans and suggests the potential for exposure to these microbes to be a health hazard.

PURPOSE: Immunotherapy has recently been shown to improve outcomes for advanced PD-L1-positive triple-negative breast cancer (TNBC) in the Impassion130 trial, leading to FDA approval of the first immune checkpoint inhibitor in combination with taxane chemotherapy. To further develop predictive biomarkers and improve therapeutic efficacy of the combination, interrogation of the tumor immune microenvironment before therapy as well as during each component of treatment is crucial. Here we use single-cell RNA sequencing (scRNA-seq) on tumor biopsies to assess immune cell changes from two patients with advanced TNBC treated in a prospective trial at predefined serial time points, before treatment, on taxane chemotherapy and on chemo-immunotherapy. METHODS: Both patients (one responder and one progressor) received the trial therapy, in cycle 1 nab-paclitaxel given as single agent, in cycle 2 nab-paclitaxel in combination with pembrolizumab. Tumor core biopsies were obtained at baseline, 3 weeks (after cycle 1, chemotherapy alone) and 6 weeks (after cycle 2, chemo-immunotherapy). Single-cell RNA sequencing (scRNA-seq) of both cancer cells and infiltrating immune cells isolated were performed from fresh tumor core biopsy specimens by 10x chromium sequencing. RESULTS: ScRNA-seq analysis showed significant baseline heterogeneity of tumor-infiltrating immune cell populations between the two patients as well as modulation of the tumor microenvironment by chemotherapy and immunotherapy. In the responding patient there was a population of PD-1(high) expressing T cells which significantly decreased after nab-paclitaxel plus pembrolizumab treatment as well as a presence of tissue-resident memory T cells (T(RM)). In contrast, tumors from the patient with rapid disease progression showed a prevalent and persistent myeloid compartment. CONCLUSIONS: Our study provides a deep cellular analysis of on-treatment changes during chemo-immunotherapy for advanced TNBC, demonstrating not only feasibility of single cell analyses on serial tumor biopsies but also the heterogeneity of TNBC and differences in on-treatment changes in responder versus progressor.

Physics teachers are often faced with the challenge of having to set up difficult experiments, or they have to consciously manipulate parameters in order to be able to demonstrate a phenomenon convincingly. Comprehensive laboratory courses are standard procedure in any study program for prospective physics teachers. However, many students, even after completing standard laboratory courses, show difficulties in standard experimental situations, such as measuring an electric current. We report on a new seminar concept for students in physics teacher study programs. This concept is based on the current state of research in physics education, on the teachers’ professional competences, and on the modelling of experimental competence, justifying its embedding in teacher training. We present first results of a pilot study carried out to evaluate the seminar.

Rationale: In lung cancer, upregulation of the PI3K (phosphoinositide 3-kinase) pathway is an early event that contributes to cell proliferation, survival, and tissue invasion. Upregulation of this pathway was recently described as associated with enrichment of the lower airways with bacteria identified as oral commensals. Objectives: We hypothesize that host–microbe interactions in the lower airways of subjects with lung cancer affect known cancer pathways. Methods: Airway brushings were collected prospectively from subjects with lung nodules at time of diagnostic bronchoscopy, including 39 subjects with final lung cancer diagnoses and 36 subjects with noncancer diagnoses. In addition, samples from 10 healthy control subjects were included. 16S ribosomal RNA gene amplicon sequencing and paired transcriptome sequencing were performed on all airway samples. In addition, an in vitro model with airway epithelial cells exposed to bacteria/bacterial products was performed. Measurements and Main Results: The composition of the lower airway transcriptome in the patients with cancer was significantly different from the control subjects, which included up-regulation of ERK (extracellular signal–regulated kinase) and PI3K signaling pathways. The lower airways of patients with lung cancer were enriched for oral taxa (Streptococcus and Veillonella), which was associated with up-regulation of the ERK and PI3K signaling pathways. In vitro exposure of airway epithelial cells to Veillonella, Prevotella, and Streptococcus led to upregulation of these same signaling pathways. Conclusions: The data presented here show that several transcriptomic signatures previously identified as relevant to lung cancer pathogenesis are associated with enrichment of the lower airway microbiota with oral commensals.

Mortality among patients with COVID-19 and respiratory failure is high and there are no known lower airway biomarkers that predict clinical outcome. We investigated whether bacterial respiratory infections and viral load were associated with poor clinical outcome and host immune tone. We obtained bacterial and fungal culture data from 589 critically ill subjects with COVID-19 requiring mechanical ventilation. On a subset of the subjects that underwent bronchoscopy, we also quantified SARS-CoV-2 viral load, analyzed the microbiome of the lower airways by metagenome and metatranscriptome analyses and profiled the host immune response. We found that isolation of a hospital-acquired respiratory pathogen was not associated with fatal outcome. However, poor clinical outcome was associated with enrichment of the lower airway microbiota with an oral commensal (Mycoplasma salivarium), while high SARS-CoV-2 viral burden, poor anti-SARS-CoV-2 antibody response, together with a unique host transcriptome profile of the lower airways were most predictive of mortality. Collectively, these data support the hypothesis that 1) the extent of viral infectivity drives mortality in severe COVID-19, and therefore 2) clinical management strategies targeting viral replication and host responses to SARS-CoV-2 should be prioritized.

In lung cancer, enrichment of the lower airway microbiota with oral commensals commonly occurs, and ex vivo models support that some of these bacteria can trigger host transcriptomic signatures associated with carcinogenesis. Here, we show that this lower airway dysbiotic signature was more prevalent in the stage IIIB–IV tumor–node–metastasis lung cancer group and is associated with poor prognosis, as shown by decreased survival among subjects with early-stage disease (I–IIIA) and worse tumor progression as measured by RECIST scores among subjects with stage IIIB–IV disease. In addition, this lower airway microbiota signature was associated with upregulation of the IL17, PI3K, MAPK, and ERK pathways in airway transcriptome, and we identified Veillonella parvula as the most abundant taxon driving this association. In a KP lung cancer model, lower airway dysbiosis with V. parvula led to decreased survival, increased tumor burden, IL17 inflammatory phenotype, and activation of checkpoint inhibitor markers. Significance: Multiple lines of investigation have shown that the gut microbiota affects host immune response to immunotherapy in cancer. Here, we support that the local airway microbiota modulates the host immune tone in lung cancer, affecting tumor progression and prognosis. See related commentary by Zitvogel and Kroemer, p. 224. This article is highlighted in the In This Issue feature, p. 211

Resistance to targeted therapies is an important clinical problem in HER2-positive (HER2+) breast cancer. “Drug-tolerant persisters” (DTP), a subpopulation of cancer cells that survive via reversible, nongenetic mechanisms, are implicated in resistance to tyrosine kinase inhibitors (TKI) in other malignancies, but DTPs following HER2 TKI exposure have not been well characterized. We found that HER2 TKIs evoke DTPs with a luminal-like or a mesenchymal-like transcriptome. Lentiviral barcoding/single-cell RNA sequencing reveals that HER2+ breast cancer cells cycle stochastically through a “pre-DTP” state, characterized by a G0-like expression signature and enriched for diapause and/or senescence genes. Trajectory analysis/cell sorting shows that pre-DTPs preferentially yield DTPs upon HER2 TKI exposure. Cells with similar transcriptomes are present in HER2+ breast tumors and are associated with poor TKI response. Finally, biochemical experiments indicate that luminal-like DTPs survive via estrogen receptor–dependent induction of SGK3, leading to rewiring of the PI3K/AKT/mTORC1 pathway to enable AKT-independent mTORC1 activation. Significance: DTPs are implicated in resistance to anticancer therapies, but their ontogeny and vulnerabilities remain unclear. We find that HER2 TKI-DTPs emerge from stochastically arising primed cells (“pre-DTPs”) that engage either of two distinct transcriptional programs upon TKI exposure. Our results provide new insights into DTP ontogeny and potential therapeutic vulnerabilities. This article is highlighted in the In This Issue feature, p. 873

Effective public response to a pandemic relies upon accurate measurement of the extent and dynamics of an outbreak. Viral genome sequencing has emerged as a powerful approach to link seemingly unrelated cases, and large-scale sequencing surveillance can inform on critical epidemiological parameters. Here, we report the analysis of 236 SARS-CoV2 sequences from cases in the New York City metropolitan area during the initial stages of the 2020 COVID-19 outbreak. The majority of cases throughout the region had no recent travel history or known exposure, and genetically linked cases were spread throughout the region. Comparison to global viral sequences showed that the majority were most related to cases from Europe. Our data are consistent with numerous seed transmissions from multiple sources and a prolonged period of unrecognized community spreading. This work highlights the complementary role of real-time genomic surveillance in addition to traditional epidemiological indicators.

No consensus has been reached so far concerning the key topics on quantum physics suitable for secondary school teaching, despite comprehensive research. We identified the the variety of associations with quantum physics among professionals as a potential cause. From an explorative mind map study with N = 29 physics researchers, we derive six subject-specific focal points in the associations that researchers have with quantum physics.

In this article, an approach to integrate contemporary quantum physics into secondary school teaching is presented. The Erlanger concept on quantum optics provides an experimental-based guideway to aspects of modern quantum physics. We avoid the traditional historical approach in order to overcome the lack of modern concepts of quantum physics. In an acceptance survey, initial empirical evidence for the acceptance of the developed explanatory approaches was evaluated.

The electrical circuit of the jumping ring experiment based on discharging a capacitor is optimized. The setup is scoop proof at 46?V and yet the ring jumps more than 9?m high. The setup is suitable for both lectures and student laboratory work in higher education.

A high standard of living causes a large number of irreversible processes, which generate entropy. Energy flow is necessary to remove entropy from the location of generation; commonly this is called energy consumption. The usefulness of an energy flow is defined by its capability of removing entropy from an irreversible process. Fossil energy sources are limited and their use causes ecological damage and conflicts about distribution. As a long-term alternative, energy needs to be obtained from the incoming stream of solar radiation, or indirectly by replacing natural heat engines with artificial ones. Due to the environmental side effects of this replacement, unnecessary artificial entropy production has to be minimized.

An ultra-capacitor in parallel with the output of a dc power supply reduces the source resistance and yields an electrical current of up to 900A through a low resistance load. The magnetic field of wires is made visible with iron filings over a large area.

We present an experiment for education which demonstrates random transmission or reflection of heralded single photons on beam splitters. With our set-up, we can realize different quantum random experiments by appropriate settings of polarization rotators. The concept of entanglement is motivated by correlated randomness. The experiments are suitable for undergraduate education and are available as interactive screen experiments.

Single photons are used for fundamental quantum physics experiments as well as for applications. Originally being a topic of advance courses, such experiments are increasingly a subject of undergraduate courses. We provide interactive screen experiments (ISE) for supporting the work in a real laboratory, and for students who do not have access to a quantum optics laboratory. The main focus of the ISE is on undergraduate education, but some of the experiments are suitable for other levels of higher education as well.

A digital SLR camera with a standard lens (50 mm focal length, f/1.4) on a fixed tripod is used to obtain photographs of the sky which contain stars up to 8m apparent magnitude. The angle of view is large enough to ensure visual identification of the photograph with a large sky region in a stellar map. The resolution is sufficient to observe the motion of Saturn, and of the satellites of Jupiter, within 24 h.

The simultaneous sound of several voices or instruments requires proper tuning to achieve consonance for certain intervals and chords. Most instruments allow enough frequency variation to enable pure tuning while being played. Keyboard instruments such as organ and piano have given frequencies for individual notes and the tuning must be based on a compromise. The equal temperament is not the only solution, but a special choice. Unequal temperaments produce better results in many cases, because important major thirds and triads are improved. Equal temperament was not propagated by Johann Sebastian Bach, as is often stated in introductory literature on this topic.

We report on an injection-seeded 9.5-W 82-MHz-repetition-rate picosecond optical parametric generator (OPG) based on a 55 mm long crystal of periodically poled lithium niobate (PPLN) with a quasi-phase-matching (QPM) grating period of 29.75 μm. The OPG is excited by a continuously diode pumped mode-locked picosecond Nd:YVO4 oscillator-amplifier system. The laser system generates 7 ps pulses with a repetition rate of 82.3 MHz and an average power of 24 W. Without injection-seeding the total average output power of the OPG is 8.9 W, which corresponds to an internal conversion efficiency of 50%. The wavelengths of the signal and idler waves were tuned in the range 1.57–1.64 μm and 3.03–3.3 μm, respectively, by changing the crystal temperature from 150 °C to 250 °C. Injection seeding of the OPG at 1.58 μm with 4 mW of single frequency continuous-wave radiation of a distributed-feedback (DFB) diode laser increases the OPG output to 9.5 W (53% conversion efficiency). The injection seeding increases the pulse duration and reduces the spectral bandwidth. When pumped by 10 W of 1.06 μm laser radiation, the duration of the signal pulses increased from 3.6 ps to 5.5 ps while the spectral bandwidth is reduced from 4.5 nm to 0.85 nm. Seeding thus improved the time-bandwidth product from 1.98 to a value of 0.56, much closer to the Fourier limit.

This paper reports on the pulse-to-pulse wavelength tuning of a multiwatt narrow-linewidth nanosecond optical parametric generator (OPG). The OPG consists of a 55 mm long periodically poled lithium niobate (PPLN) crystal with a domain period of 29.75 μm. Excited by 5.7 W of 1064 nm laser pulses (duration: 10 ns, repetition rate: 10 kHz, spatial beam quality M2=1.2) the OPG generates 1.78 W of signal radiation and 0.79 W of idler radiation. The total output of 2.57 W corresponds to a crystal internal efficiency of 64%. The wavelength and spectral width of the OPG is controlled by injecting the cw single mode radiation of a 1580.2 nm DFB diode laser. A seed power as low as 20 μW is sufficient for complete control of the wavelength and the (90 MHz) linewidth of the OPG output. Fast tuning of the diode laser frequency is possible with a rate of up to 6×107 GHz/s by changing the diode current. This allows pulse-to-pulse wavelength tuning of the 10 kHz OPG over a range of 25 GHz limited only by the maximum variation of the current (of 60 mA) allowed for the DFB diode laser used. Increasing the current from 35 mA to 95 mA changed the diode power from 0.3 mW to 5.7 mW. Since these power levels are well above the required minimum seed power, the change of the diode current changes the wavelength of the OPG, but not its linewidth or its output power.

We explore the potential of a new laser resonator design that uses an intracavity Fourier transformation and allows for multi-color operation, gain extension and intracavity second harmonic generation (SHG). First, purely electronically controlled wavelength tuning is demonstrated using liquid crystal displays (LCDs) and digital mirror devices. Moreover, the new design can be applied to different lasers as shown on the examples of a laser diode and a Thulium-doped fiber laser in the mid-infrared. Furthermore, we demonstrate the simultaneous control of multiple gain media within one single external cavity. In addition, tunable emission in the 490 nm range is obtained using intracavity SHG with one single control parameter. Finally, we unambiguously prove simultaneous multi-wavelength operation with variable wavelength spacings and apply it to the generation of THz difference frequencies.

The Barkhausen effect can be impaired by various noise sources. These are avoided by a hum-bucking coil, earthing, a preamplifier with carefully chosen bandwidth, and matching the input resistor to the inductance of the coil. The present setup is suitable for lecture hall demonstration in higher education institutes.

The discovery and potency optimization of a series of 7-aminofuro[2,3-c]pyridine inhibitors of TAK1 is described. Micromolar hits taken from high-throughput screening were optimized for biochemical and cellular mechanistic potency to ∼10 nM, as exemplified by compound 12az. Application of structure-based drug design aided by co-crystal structures of TAK1 with inhibitors significantly shortened the number of iterations required for the optimization.

Acceleration of a common car with with a turbocharged diesel engine is compared to the same type with an electric motor in terms of kinematics. Starting from a state of rest, the electric car reaches a distant spot earlier than the diesel car, even though the latter has a better specification for engine power and average acceleration from 0 to 100 km h−1. A three phase model of acceleration as a function of time fits the data of the electric car accurately. The first phase is a quadratic growth of acceleration in time. It is shown that the tenfold higher coefficient for the first phase accounts for most of the kinematic advantage of the electric car.

We have established a long-term culture system to study macrophages chronically infected with mycobacteria. Monocytes are infected with Bacillus Calmette-Guerin (BCG) and support exponential intracellular replication without profound perturbation of normal host cell function. We have used this system to investigate lymphokine-activated killer (LAK)-mediated cytolysis. We have found that interleukin 2 stimulation of peripheral blood lymphocytes generates a cytotoxic activity against human monocytes. A CD56- subpopulation of LAK cells specifically recognizes and lyses BCG-infected cells. Lysis of the host cell has no effect on parasite viability and results in the liberation of bacteria capable of infecting more cells.

13 patients with human immunodeficiency virus type 1 infection class II-IV, but without opportunistic infection or neoplasm, received 6 micrograms (3.6 x 10(4) IU) of polyethylene glycol recombinant human interleukin 2 (PEG IL-2) intradermally twice a week for 4 mo were then followed for an additional 6 mo. Clinical, immunological, and viral parameters were monitored in the patients, all of whom were taking zidovudine. The cutaneous administration of PEG IL-2 resulted in an indurated zone resembling a delayed-type hypersensitivity response of 26 +/- 1 mm diameter (676 mm2) at 72-96 h after injection throughout the 4 mo of administration. This dose, which was appreciably lower than in most previous trials, was not associated with local or systemic toxicity. No increase in the viral burden of circulating leukocytes or plasma occurred. A number of immunological functions were stimulated by this course of therapy. All patients demonstrated high levels of lymphokine-activated killer cell activity by cells freshly removed from the circulation and in the absence of in vitro exposure to IL-2. Natural killer cell activity was also enhanced. Limiting dilution analysis revealed an increase in the frequency of IL-2-responsive cells from abnormally low to levels above normal during the course of injections. In a subgroup of four patients with > or = 400 CD4+ T cells/microliter at entry, there was a trend to sustained increases in CD4+ T cell numbers. However, this increase did not reach statistical significance. This subset of patients also exhibited higher proliferative responses to phytohemagglutinin as mitogen. Several of these effects persisted for 3-6 mo after cessation of therapy. In conclusion, low-dose IL-2 regimens lead to sustained immune enhancement in the absence of toxicity. We suggest pursuit of this approach for further clinical trials both as prophylaxis and therapy.

Interleukin-2 (IL-2) is a key cytokine in cellular immunity. Human immunodeficiency virus type 1 (HIV-1)-infected individuals lack IL-2 because of low CD4+ T lymphocyte numbers. In an attempt to enhance cellular immunity, low-dose recombinant human (rh) IL-2 at 10 micrograms or 180,000 units or its polyethylene glycol (PEG) derivative at 9 micrograms or 36,000 units was given by intracutaneous injection to 8 HIV-1-infected men for 30 days. Participants had no evidence of opportunistic infection and received concurrent zidovudine. IL-2 treatment was nontoxic and elicited a local cellular response resembling classic delayed-type hypersensitivity (DTH) with local interferon-gamma production, even in anergic patients. Systemic responses included enhanced DTH responses to recall antigens, improved in vitro proliferative responses to mitogen, and enhanced NK cell activity. Peripheral leukocyte phenotype and virus titers were unchanged. Long-term studies of low-dose IL-2 are warranted to determine whether immunoenhancing effects can be sustained and if they are associated with improved clinical course.

Recombinant granulocyte/macrophage-colony-stimulating factor (rGM-CSF), prepared from Chinese hamster ovary (CHO) cells and Escherichia coli, was administered to 35 patients with the borderline and polar lepromatous forms of leprosy by the intradermal and subcutaneous routes at doses of 7.5-45.0 micrograms/d for 10 d. With each of these doses and routes, increases in the number of circulating eosinophils were noted. After the intradermal injection, the local skin sites demonstrated zones of roughening and micronodularity that appeared within 24-48 h and persisted for more than 6 d. Reinjection of sites led to enhanced areas of epidermal reaction. GM-CSF prepared from CHO cells was a more potent inducer of this effect. GM-CSF given by the subcutaneous route, at higher doses, failed to initiate these changes. At the microscopic level, the epidermis became thickened (+75%) with increased numbers and layers of enlarged keratinocytes. These contained increased numbers of ribosomes and prominent nucleoli, and were imbedded in a looser meshwork of the zona Pellucida. The modified keratinocytes remained MHC class II antigen negative throughout the course of the response. A major change in the dermis was the progressive accumulation of CD1+, Birbeck granule-positive cells. These Langerhans were recognizable at 48 h after intradermal injection and reached maximum numbers by 4 d. During this period the number of epidermal Langerhans cells remained relatively constant. No increment in dermal Langerhans cells occurred when GLM-CSF was injected by the subcutaneous route. No appreciable increase in the numbers of T cells and monocytes was noted, and granulocytes and eosinophils were largely present within the dermal microvasculature. 4-mm punch biopsies taken from injected sites and adjacent controls were compared in terms of the rapidity of wound healing. 22 of 26 sites demonstrated more rapid filling and hemostasis, whereas four were equivalent to controls. We conclude that rGM-CSF, when introduced into the skin, leads to enhanced keratinocyte growth, the selective recruitment of Langerhans cells into the dermis, and enhanced wound healing of the prepared site. There was no evidence of an enhanced cell-mediated response to Mycobacterium leprae, and bacillary numbers remained unchanged.

We report on the observation of self-injection-locking of the signal wave of an optical parametric oscillator (OPO) with the intracavity frequency doubled idler wave. The two-mirror OPO is based on a periodically poled LiNbO3 (PPLN) crystal and pumped with a grating stabilized, continuous-wave (CW) single-frequency diode master-oscillator power-amplifier (MOPA) system. Simultaneous quasi-phase-matching (QPM) of OPO and second harmonic generation (SHG) is provided in the same crystal which carries two different domain gratings. The beat of the signal wave and the frequency-doubled idler wave is suppressed within a 500-kHz wide frequency range centered around zero as expected for self-injection- locking. The measurements prove the feasibility of optically phase-stabilized by-three-division of an optical frequency with CW-OPOs using cascaded nonlinearities.

We report on a diode-laser pumped cw optical parametric oscillator (OPO) based on quasi-phase-matched periodically poled lithium tantalate. Pumped by the 2.3-W single-frequency, nearly diffraction-limited 925-nm output of an InGaAs diode master-oscillator power amplifier, the pump and signal resonant OPO generates a single-frequency idler wave with an output of as much as 244 mW. The wavelengths of the signal and idler waves are widely tunable in the range 1.55-2.3mum by use of different poling periods (27.3 to 27.9mum) and by variation of the crystal temperature in the range 70-190 degrees C.

We describe a cw optical parametric oscillator (OPO) with multigrating periodically poled LiTaO(3) . Pumped by a single-frequency 532-nm laser, the OPO emits single-frequency radiation at wavelengths from 0.77 to 1.73 microm with as much as 60 mW of output power. Mode-hop-free operation for as long as 50 min, a low frequency drift (70 MHz/h), and as much as 700-MHz continuous frequency tuning of signal and idler are demonstrated.

We report on what is believed to be the first singly resonant cw optical parametric oscillator (SRO) that is directly pumped by a diode laser. The SRO consists of a 38-mm-long periodically poled LiNbO(3) crystal in a four-mirror signal-resonant ring cavity. Pumped by 2.5 W of 925-nm diode-laser radiation, the SRO generates 480 mW of single-frequency idler radiation at 2.1mum . The wavelengths of the signal and the idler output are tuned in the ranges of 1.55 to 1.70mum and 2.03 to 2.29mum, respectively, by tuning the wavelength of the diode laser from 924.0 to 925.4 nm.

We demonstrate the generation of sub-6-fs pulses centered at 405 nm by frequency doubling of 8.6-fs Ti:sapphire laser pulses. The frequency doubling is carried out in a nonlinearly chirped quasi-phase-matching grating fabricated in a lithium tantalate substrate. This device simultaneously provides frequency conversion and pulse compression of the positively prechirped fundamental pulses. The second-harmonic pulses are characterized in a cross-correlation setup, and their pulse shapes are retrieved by two iterative phase-reconstruction algorithms. The generated second-harmonic spectrum spans a bandwidth of 220 THz. To our knowledge, these are the shortest pulses ever generated in the blue spectral region.

A rotor of 12 cm diameter is attached to a precision electric motor, used as a generator, to make a model wind turbine. Output power of the generator is measured in a wind tunnel with up to 15 m s−1 air velocity. The maximum power is 3.4 W, the power conversion factor from kinetic to electric energy is cp = 0.15. The v3 power law is confirmed. The model illustrates several technically important features of industrial wind turbines quantitatively.