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Recent immunoelectron microscopic studies have shown that immunoreactive prolactin (PRL) in rat pituitary can be detected not only in typical PRL cells, characterized by large secretory granules, but also in another type of cell, which contains small secretory granules. To determine whether or not these two cell types are involved in PRL biosynthesis, we developed a procedure to investigate PRL gene expression by using in situ hybridization at the ultrastructural level. Rat pituitary was fixed and vibratome sections were incubated with a PRL [35S]-cDNA probe and subsequently flat-embedded in Araldite. Semi-thin and ultra-thin sections were processed for autoradiography. The results indicate that only the two PRL cell types were labeled. When immunolabeling for PRL was applied to ultra-thin sections, only immunopositive cells were seen to contain silver grains. In these cells the silver grains were associated with the rough endoplasmic reticulum and nucleus. When a growth hormone (GH) [35S]-cDNA probe was used as a control, only GH-secreting cells were labeled. This study confirms that the two PRL cell types are involved in biosynthesis of PRL. Moreover, this simple in situ hybridization technique provides a new approach to accurately localize mRNA in complex tissue and to investigate the subcellular distribution of mRNA under differing experimental conditions.

The elastic-plastic-creep characteristics of solder joints are implemented in a nonlinear finite element program ABAQUS by developing user defined material subroutines. An eutectic Pb-Sn solder joint of a resistor carrier under thermal cycling between 125°C and −55°C is modeled, and the effect of various parameters on the solder joint cycle life is evaluated. The strain range of the solder joint under thermal cycling loads is calculated, which is then converted into solder joint cycle life through a fatigue-life relationship proposed by Engelmaier (1983). The parameters studied include: ramp time, hold time, grain size, initial temperature, constitutive equations, material properties for solder alloys, and mesh refinement. The effects of these variations on the fatigue life of solder joints are illustrated. The described method can serve as a tool in the design and manufacturing of surface-mount (SMT) assemblies.

The perception of temporal modulations of pulsatile electric stimuli was measured in seven cochlear implant patients using the Cochlear Pty. Limited prosthesis. Four patients were postlingually deafened adults and three patients were young adults who were deafened very early in life. The first study measured detection thresholds for modulated pulse duration for a series of modulation frequencies and pulse rates. The shape of the detection thresholds as a function of modulation frequency, the temporal modulation transfer function, often resembled a low‐pass filter with a 50–100‐Hz cut‐off frequency. Thresholds did not markedly vary across the different pulse rates for most patients. Thresholds were less than 10%–20% of the range of usable hearing for most patients. The second study compared detection thresholds for modulated pulse durations around different reference pulse durations: 50, 100, and 300 μs. Detection thresholds were generally proportional to the different reference pulse durations. The third study measured difference limens for the discrimination of modulation depth. The difference limens were similar to the detection thresholds for the same reference pulse duration and pulse rate. The three patients deafened very early in life showed more within‐group variation in performance, and their overall levels of performance were poorer than those of the postlingually deafened adults.

PP2C is a structurally diversified protein phosphatase family with a wide range of functions in cellular signal transduction. A novel PP2C subtype, designated PP2Cdelta, was identified from a rat cDNA clone, which encodes a protein of 392 amino acid residues. While PP2Cdelta shares approximately 30% sequence identity in its catalytic domain with the mammalian PP2C, it lacks a 90-residue carboxyl-terminal sequence conserved in mammalian PP2C. Northern blot analysis showed that PP2Cdelta is widely expressed in rat tissues. The transcription of the PP2Cdelta gene was activated in response to stress, such as the addition of ethanol to the culture medium or UV irradiation of cells. Recombinant PP2Cdelta purified from bacteria exhibited a potent Mn2+-dependent serine/threonine phosphatase activity. Unlike other members of the PP2C family, the activity of PP2Cdelta was inhibited, rather than stimulated, by Mg2+. Transfection with PP2Cdelta resulted in inhibition of cell growth, precluding generation of stable 293 or CHO transfectants. Using a modified tetracycline-regulated PP2Cdelta-GFP dicistronic expression cassette, it was revealed that overexpression of PP2Cdelta blocked cell cycle progression and arrested cells at early S phase, resulting in inhibition of DNA synthesis and leading to cell death. These results suggest that PP2Cdelta plays a role in regulation of cell cycle progression via dephosphorylation of its substrates whose appropriate phosphorylation states might be crucial for cell proliferation.

1 Contractile responses of smooth muscle from the Wistar rat urinary bladder were studied with the use of muscarinic agonists and antagonists. 2 McN-A-343 induced only weak contractile responses of the bladder muscle. In contrast, oxotremorine showed higher potency than either acetylcholine or bethanechol in inducing a contractile response (the respective pD2 values were 6.38 ± 0.25, 4.82 ± 0.24 and 4.42 ± 0.14). 3 The M2 antagonists, methoctramine (10⁻⁹ m to 10⁻⁵ m) and gallamine (10⁻⁹ m to 10⁻⁵ m), did not reduce acetylcholine-induced (10⁻⁵ m) contractions of the bladder muscle strip. On the other hand, 4-diphenyl-acetoxy-N-methyl piperidine methiodide (4-DAMP, 10⁻¹⁰ m to 10⁻⁷ m), an M3 receptor blocker, effectively antagonized the acetylcholine-induced contractions in a concentration-dependent manner. 4-DAMP had a similar pA2 value to those of the non-selective antagonists, atropine and scopolamine (pA2 values were 8.26 ± 0.05, 8.36 ± 0.05 and 8.41 ± 0.11, respectively). Pirenzepine, an M1 blocker, antagonized the contractions at higher concentrations (10⁻⁸ m to 10⁻⁵ m, pA2 = 6.23 ± 0.04). 4 It is concluded that (1) the dominant muscarinic receptor subtype responsible for smooth muscle contraction in the rat urinary bladder is M3; and (2) the muscarinic agonist oxotremorine was more potent than acetylcholine and bethanechol in inducing a contractile response.

The heterogeneization of the catalytic biphasic system Pd(OAc)2/5tppts/water/nitrile (tppts = sodium salt of tri(m-sulphophenyl)phosphine)) is performed by controlled deposition on silica of the catalyst in its aqueous phase (SAP catalyst). The resulting heterogeneous molecular catalysts are active for the alkylation of ethyl acetoacetate or morpholine by (E)-cinnamyl ethyl carbonate. They are very selective for the monoalkylated product, and the secondary reactions due to water itself, such as formation of cinnamyl alcohol, are suppressed. Finally, the active Pd(0) entity is stable toward decomposition into metallic particles. This method (SAP) is most appropriate with true biphasic media. Thus, with CH3CN, the SAP catalyst is less active than the monophasic homogeneous catalyst, whatever the water content of the solid. On the contrary, with PhCN, the SAP catalyst is drastically more active than the biphasic homogeneous catalyst, a phenomenon which must be correlated to a large enhancement of the interphase surface area. The water content of the SAP catalyst intervenes in different ways. Thus, a minimum amount of water (ca. 15−20 wt %) is necessary to observe any catalytic activity, a phenomenon which is correlated with a certain mobility of the complex on the surface of silica, confirmed by ³¹P MAS NMR, but also probably to a minimum concentration of the reactants in the aqueous phase. Activity increases when the water content reaches 30−50 wt %. Within these boundaries, no detectable palladium leaching is observed but the catalysts progressively loose part of their activity after several recyclings, a phenomenon which seems to be correlated with water leaching into the organic phase. ³¹P MAS NMR reveals that the main surface complex is Pd(0)(tppts)3. Its chemical shift corresponds to the weighted average of that of Pd(tppts)3 (δ 22.6 ppm) and that of free tppts (δ −5.6 ppm), in accordance with a fast exchange between free tppts and coordinated tppts. No strong interaction between the complex and the silica surface could be evidenced; in the presence of small amounts of water, free tppts interacts with the silica surface (δ ≈ 0 ppm) via hydrogen bonding. Above 50 wt % of water, a number of problems are encountered which lead to an overall drop in activity:  the solid agglomerates and palladium and water leaching both become significant. The SAP catalysts supported on a nonporous silica present the same behavior, i.e., increasing activity with increasing water content up to a content corresponding roughly to the wetting volume.

We have characterised an etoposide-resistant subline of the small-cell lung cancer cell line, UMCC-1, derived at our centre. Subline UMCC-1/VP was developed by culturing the parent line in increasing concentrations of etoposide over 16 months. UMCC-1/VP is 20-fold resistant to etoposide by MTT assays, relative to the parent line, and is cross-resistant to doxorubicin, vincristine and actinomycin D, but not to taxol, cisplatin, melphalan, thiotepa or idarubicin. Topoisomerase II immunoblotting demonstrates a 50% reduction of the protein in the resistant subline. The UMCC-1/VP subline demonstrates a marked decrease in the accumulation of [3H]etoposide relative to the parent line, as well as a modest reduction in the accumulation of daunorubicin. Reverse transcription-polymerase chain reaction assays demonstrate no detectable mdr1 expression but marked expression of the multidrug resistance-associated protein (MRP) gene in the resistant subline. Northern blotting with an MRP cDNA probe confirms marked overexpression of the MRP gene only in the UMCC-1/VP subline. Western blotting with antisera against MRP peptide confirms a 195 kDa protein band in the UMCC-1/VP subline. Southern blotting experiments demonstrate a 10-fold amplification of the MRP gene in the resistant subline. Depletion of glutathione with buthionine sulphoximine sensitised UMCC-1/VP cells to daunorubicin and etoposide. Our studies indicate that MRP gene expression may be induced by etoposide and may lead to reduced accumulation of the drug.

Abstract. A monoclonal antibody was used for the localization of the nuclear T3receptor in different tissues of the adult rat: the liver, kidney, heart, lung, spleen, testis, and pituitary. In the liver, the immunoreactivity was found uniformly distributed in the nuclei of hepatocytes. Sections incubated with a control ascitic fluid or with the same ascitic fluid pre-adsorbed with purified receptor showed no specific staining. In the kidney, the immunoreactivity was higher in the epithelial cell of the proximal convoluted tubes and juxtaglomerular cells. In the heart, only the myocardial cells were stained. In the lung, the immunoreactivity was confined to type II pneumocytes and alveolar macrophages. In the spleen, only a few mature lymphocyte and macrophage cell nuclei were stained. These results show that: 1) the abundance of the nuclear T3correlates with previous studies using hormone binding techniques; 2) the nuclear T3receptor is selectively located in certain cell types, which possess a precise local function.

Majority systems are encountered in both decision theory and reliability theory. In decision theory for example a jury or committee employing a majority rule will make the ‘correct' decision if a majority of the individuals do so. In reliability theory some coherent systems function if and only if a majority of the components work properly. In this paper results concerning the reliability of majority systems are developed which are applicable in both areas. Two models incorporating dependence between individuals or components in majority systems are introduced, and various monotonicity results for their reliability functions are established. Comparisons are also made between direct (or simple) and indirect majority systems.

Wavelength tuneable pulses of about 100fs duration have been generated through self-mode-locking initiated by synchronous pumping of a c.w. Cr4+ : YAG laser.

Discharge patterns of single auditory nerve fibers were recorded from normal-hearing cats implanted with a 12-band intracochlear electrode array. Stimuli were biphasic current pulses of specifiable width, amplitude, and rate. Acoustic tuning curves were obtained to determine the cochlear positions of the fibers. Response latencies to electrical stimuli formed two groups. Short latency (0.3 to 0.7 ms) responses were attributed to direct activation of spiral ganglion neurons. At high stimulus intensities, these often exhibited abrupt shifts toward even shorter latencies. Long latency (> 1.5 ms) responses were probably caused by electrophonic activation of functional hair cells. Response thresholds to electrical stimuli depended on a fiber's proximity to the stimulating electrodes, and they did not depend on a fiber's acoustic response threshold or spontaneous discharge rate. High intensity (> 1.5 mA) stimuli could excite fibers over a wide range of characteristic frequencies, even for the narrowest (0.45 mm) electrode separations. Response threshold was an exponentially decreasing function of pulse width for widths up to 300 μs/phase. Fiber discharges were highly phase-locked at all suprathreshold intensities, and saturation discharge rates usually equaled stimulus pulse rates for rates up to at least 800 pulses/s. Dynamic ranges were small (1 to 6 dB), increased with pulse rate, and were uncorrelated with electrical response threshold. Within the dynamic range, shapes of poststimulus time and interspike interval histograms resembled those obtained in response to acoustic stimuli. Depolarization block caused fiber activity to cease in 2 to 5 seconds for sustained stimuli presented at high (> 600 pulses/s) pulse rates and intensities.

The theory of majorization is applied to investigate the effects of heterogeneity in a class of epidemics. In particular, the heterogeneity of the components of the contact vector of a given susceptible is studied. It is shown that if the heterogeneity can be partially ordered through majorization, then the probability functions of escaping infection can be similarly ordered. The results can be applied to AIDS research on the spread of the HIV virus.

A consonant confusion study was undertaken on a multiple-channel cochlear implant patient using a wearable speech processing device. This patient suffered from total bilateral deafness acquired postlingually. The consonants/b/,/p/,/m/,/v/,/f/,/d,/t/,/n/,/z/,/s/,/g/,/ld were presented in a VCV context with the vowel /a/ as in fatherby a male and female speaker trader three conditions: lipreading alone; electrical stimulation alone using the wearable speech processor and multiple-channel cochlear implant; lipreading in conjunction with electrical stimulation. No significant difference was detected between the results for the male and female speakers. The percentage correct scores for the pooled results of both speakers were lipreading alone-30%; electrical stimulation alone—48%; lipreading with electrical stimulation-70%. Performance was significantly better for lipreading with electrical stimulation than for lipreading alone and for electrical stimulation alone than for lipreading alone. An information transmission analysis demonstrated the effective integration of visual and auditory information for lipreading with electrical stimulation. There was a significant improvement in performance for the electrical stimulation alone condition over the 2 months of the study in contrast to no such improvement for lipreading alone.

A correspondence between Hermitian modular forms and vector valued harmonic forms in locally symmetric spaces associated to is constructed and also shown in general to be nonzero. The construction utilizes Rallis-Schiffmann type theta functions and simplified arguments to circumvent differential geometric calculations used previously in related problems.

A 22-electrode implantable hearing prosthesis uses a wearable speech processor which estimates three speech signal parameters. These are voice pitch, second formant frequency and flattened spectrum amplitude. The signal is monitored continuously for periodicity in the range 80400 Hz and, if this is present, stimulation occurs at the same rate. Otherwise, as in the case of unvoiced sounds, it occurs at the random rate of fluctuation of the signal envelope. The second formant is obtained by filtering to extract the dominant peak in the midband region and by continuous measurement of the zero crossing rate. The amplitude measured is that of the whole speech spectrum, pre-emphasized by differentiation. The values that are presented to the patient are the parameter estimates immediately prior to the stimulation pulse. Second formant frequency is coded by selection of an appropriate electrode in the cochlea and amplitude by a suitably controlled current. Automatic gain control is used to keep the dynamic range of the amplitude estimate within the 30 dB range of the circuitry.

A multi-channel cochlear implant hearing prosthesis providing 22 separate channels of stimulation has been developed. The electronics for the implantable receiver-stimulator have been incorporated on a single chip, using digital circuits and employing CMOS technology. The chip is enclosed in a titanium capsule with platinum/ceramic electrode feed-throughs. A pocket-sized speech processor and directional microphone extract the following speech parameters: signal amplitude, fundamental frequency and formant frequency. The fundamental frequency is coded as electric pulse rate, and formant frequency by electrode position. The speech processor has been realized using hybrid circuits and CMOS gate arrays.The multi-channel prosthesis has undergone a clinical trial on four postlingually deaf patients with profound-total hearing losses. The speech perception results indicate that they were able to obtain open-set speech recognition scores for phonetically balanced words, CID sentences and spondees. In all cases the tests showed significant improvements when using the cochlear prosthesis combined with lipreading compared to lipreading alone.

Further improvements in speech perception for cochlear implant patients in quiet and in noise should be possible with speech processing strategies using binaural implants. For this reason, presented here is a series of initial psychophysical and speech perception studies on the authors’ first binaural cochlear implant patient. For an approximate matching of the places of stimulation on the two sides, the patient usually reported a single percept when the two sides were simultaneously stimulated. Lateralization was strongly influenced by amplitude differences between the electrical stimuli on the two sides, but only weakly by interaural time delays. Speech testing, comparing monaural with binaural electrical stimulation, showed a binaural advantage particularly in noise.

Expression of human parathyroid hormone receptor (hPTHR) was obtained in Xenopus oocytes. Receptor function was detected by hormone stimulation of endogenous Ca2+-activated Cl- current. This current was blocked by injected, but not by extracellular, EGTA, confirming that the hPTHR activates cytosolic Ca2+ signaling pathways. PTH responses were acutely desensitized but were regained in 6 12 h. Injection of cAMP or analogues had no effect on either responsiveness or desensitization to hPTH. The hPTH response was more sluggish than seen with serotonin 5-hydroxytryptamine (5-HT2C) receptor. In oocytes co-expressing both hPTHR and 5-HT2C receptors, homologous desensitization was seen, but cross-desensitization was not observed. Injection of inositol 1,4,5-trisphosphate (InsP3) elicited a fast inward current similar to that induced by serotonin, and complete cross-desensitization occurred between the InsP3 and 5-HT2C responses. Desensitization by hPTH did not affect responses to either InsP3 or serotonin, but cells desensitized to injected InsP3 still responded strongly to PTH. Oocytes did not respond to either cADPR or NAADP+, but NADP+ and analogues were found to be potent inhibitors of PTH signaling. We suggest that PTH cytosolic Ca2+ signaling in oocytes either involves a novel signaling system or proceeds through a Ca2+ compartment whose responsiveness is regulated in a novel way.

The tumor suppressor gene APC was recently identified, and the cDNA was cloned from chromosome 5q21. Point mutations affecting APC are seen in the hereditary syndrome familial adenomatous polyposis, and point mutations in APC and a closely linked gene, MCC, as well as loss of heterozygosity involving chromosome 5q have been reported in sporadic colon cancer. To our knowledge, loss of heterozygosity involving APC or MCC or both has not yet been described in any other human cancer besides lung cancer. We used the polymerase chain reaction and DNA content flow cytometric nuclear sorting to examine 30 primary human esophageal cancers for loss of heterozygosity of APC or MCC or both. Loss of one allele was detected in 77% of 26 informative cases. These data suggest that loss of heterozygosity of regions on 5q including the APC and MCC genetic loci is involved in the development and/or progression of most human esophageal cancers. They imply that inactivation of APC, MCC, and/or a linked gene on chromosome 5q plays a role in the pathogenesis of some cancers of the upper gastrointestinal tract, as well as in colon cancer and familial adenomatous polyposis.

Within the frontier orbital picture for metal chemistry, the local density of states at the Fermi energy (Ef-LDOS) plays the equivalencies of the highest-occupied molecular orbitals (HOMO) and the lowest-unoccupied molecular orbitals (LUMO) in the molecular cases. We show here that one can effectively engineer the clean surface Ef-LDOS, i.e., the frontier orbitals, of a platinum catalyst already formed within a zeolite−NaY matrix, by simply exchanging the counterion in the matrix. The potassium (K) increases while the proton (H) decreases the clean surface Ef-LDOS. The degree of enhancement or reduction depends on the degree of ion-exchange. When the intramolecular stretching frequencies of chemisorbed carbon monoxide (CO) on these Ef-LDOS enhanced and decreased samples were measured, the values extrapolated to zero coverage show an almost linear relationship with their corresponding clean surface Ef-LDOS before chemisorption.

1. β-Phenylpropionylthiocholine and N-(5-aminopentyl)-5-dimethylaminonaphthalene-1-sulphonamide (dansylcadaverine) serve as a pair of water-soluble (pH7.5) model substrates for transamidating enzymes. Amide formation could be followed directly through fluorescence measurements by monitoring the continuous extraction of the water-soluble coupling product, N-(β-phenylpropionyl)dansylcadaverine, into n-heptane. By this procedure, the steady-state kinetics of glutamine-lysine endo-γ-glutamyltransferase from human plasma (fibrinoligase, thrombin- and Ca2+-activated blood coagulation Factor XII) and from guinea-pig liver (liver transglutaminase) were investigated at 25 degrees C. 2. With β-phenylpropionylthiocholine as the varied substrate, Lineweaver-Burk plots with various concentrations of dansylcadaverine intercept on the horizontal axis, suggesting that formation of the acyl-enzyme is rate limiting. 3. On the basis of functional normality of active sites, kcat. values of 1.8 s(-1) and 0.9 s(-1) were obtained for the plasma and liver γ-glutamyltransferase respectively. The two enzymes show identical affinities for the first substrate, β-phenylpropionylthiocholine, with Ka 4 times 10(-4) M. 4. Utilization of the second substrate, dansylcadaverine, appears to be an order of magnitude more efficient with the liver enzyme. 5. N-(5-Amino-3-thiapentyl)-5-dimethylaminonaphthalene-1-sulphonamide (dansylthiacadaverine) could be used instead of dansylcadaverine in the fluorescent kinetic system. 6. Competitive inhibition by a non-fluorescent amine substrate histamine was also evaluated.

In this paper we first prove an arrangement-decreasing property of partial sums of independent random variables when they are partially ordered through the likelihood ratio ordering. We then apply a similar argument to obtain a stochastic ordering of random processes via a comparison of their parameter functions, with special applications to Poisson and Wiener processes. Finally, in Section 4 we present some applications in reliability theory, queueing, and first-passage problems.

Psychophysical studies were conducted on 10 cochlear implant patients, between 5 and 23 years of age at the time of surgery, who were deafened prior to 4 years of age. The multiple-electrode prosthesis manufactured by Cochlear Ltd. was used. Identification studies, the recognition of 2-4 stimuli after some training, were conducted on 3 of the 10 patients. For current level and repetition rate identification, performance was comparable to that observed for postlingual adult patients. For electrode position identification, however, performance was much poorer than that observed for postlingual adults. In general, the difference limens for current level, repetition rate and duration, and the gap detection thresholds were similar to those observed for postlingual adults. For 3 patients whose etiology was Usher's syndrome, the repetition rate limens at higher rates were larger than those of the other patients. The limens for electrode position, measured in a discrimination task, were 1-3 electrodes for most patients. However, for 3 patients, limens of 6-10 electrodes were recorded. For numerosity judgements, the counting of stimuli in a temporal series as a function of the rate of presentation, the patients were less successful at counting for rates of 3-8/s than for lower rates (1-2/s).

In this paper we consider a sequential version of the regression method in Monte Carlo studies under the framework of sequential estimation theory. The proposed procedure is easy to adopt on the computer, and its asymptotically optimal properties are carefully justified. Empirical results are also given.

Psychophysical and speech perception studies were conducted on eight patients using the 22‐electrode cochlear implant manufactured by Cochlear Pty. Ltd. Four early‐deafened patients became deafened at 1–3 years of age and were implanted at 5–14 years of age. Four late‐deafened (postlingual adult) patients became deafened at 38–47 years of age and were implanted at 42–68 years of age. Psychophysical studies measured the discrimination of trajectories with time‐varying electrode positions and repetition rates. Speech perception studies measured performance using two speech coding strategies: a multi‐electrode strategy which coded the first and second formant frequencies, the amplitudes of the two formants, and the fundamental frequency; and a single‐electrode strategy which coded the amplitudes of the first and second formants, and the fundamental frequency. In general, the four late‐deafened patients and one early‐deafened patient were more successful than the other three early‐deafened patients in the discrimination of electrode position trajectories and in speech perception using the multi‐electrode strategy. Three of the four late‐deafened patients were more successful than the early‐deafened patients in the discrimination of repetition rate trajectories. Speech perception performance in the single‐electrode strategy was closely related to performance in repetition rate discrimination. The improvement in speech perception performance from the single‐electrode to multi‐electrode strategy was consistent with successful performance in electrode discrimination.

Human thymidylate synthase (TS) contains three highly conserved residues Ile-108, Leu-221, and Phe-225 that have been suggested to be important for cofactor and antifolate binding. To elucidate the role of these residues and generate drug-resistant human TS mutants, 14 variants with multiple substitutions of these three hydrophobic residues were created by site-directed mutagenesis and transfected into mouse TS-negative cells for complementation assays and cytotoxicity studies, and the mutant proteins expressed and characterized. The I108A mutant confers resistance to raltitrexed and Thymitaq with respective IC50 values 54- and 80-fold greater than wild-type but less resistance to BW1843U89 (6-fold). The F225W mutant displays resistance to BW1843U89 (17-fold increase in IC50 values), but no resistance to raltitrexed and Thymitaq. It also confers 8-fold resistance to fluorodeoxyuridine. Both the kinetic characterization of the altered enzymes and formation of antifolate-resistant colonies in mouse bone marrow cells that express mutant TS are in accord with the IC50 values for cytotoxicity noted above. The human TS mutants (I108A and F225W), by virtue of their desirable properties, including good catalytic function and resistance to antifolate TS inhibitors, confirm the importance of amino acid residues Ile-108 and Phe-225 in the binding of folate and its analogues. These novel mutants may be useful for gene transfer experiments to protect hematopoietic progenitor cells from the toxic effects of these drugs.

The interesting three-dimensional hydrogen-bonded self-assembled network of [(CH2)6N4CH3]+ cations and [CoCl4]2– anions has been prepared and structurally characterized. In the title complex, the quaternization of one hexamethylenetetramine nitrogen atom has been trapped, and further stabilized by the large [CoCl4]2– anions, featuring C–HCl hydrogen bonds (3.497–3.709 ) between the methylene groups of [(CH2)6N4CH3]+ cations and the chlorine atoms of the [CoCl4]2– anions.